Inhibitors of bruton&#39;s tyrosine kinase

ABSTRACT

This application discloses compounds according to generic Formula (I): wherein all variables are defined as described herein, which inhibit Btk. The compounds disclosed herein are useful to modulate the activity of Btk and treat diseases associated with excessive Btk activity. The compounds are useful for the treatment of oncological, auto-immune, and inflammatory diseases caused by aberrant B-cell activation. Also disclosed are compositions containing compounds of Formula (I) and at least one carrier, diluent or excipient.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a continuation application of U.S. patentapplication Ser. No. 15/102,315, filed Jun. 7, 2016, which is a NationalStage Application of PCT/EP2014/077113, filed Dec. 10, 2014, whichclaims priority from U.S. Provisional Patent Application No. 61/915,576,filed Dec. 13, 2013.

FIELD OF THE INVENTION

The present invention relates to the use of novel compounds whichinhibit Btk and are useful for the treatment of oncological,auto-immune, and inflammatory diseases caused by aberrant B-cellactivation.

BACKGROUND OF THE INVENTION

Protein kinases constitute one of the largest families of human enzymesand regulate many different signaling processes by adding phosphategroups to proteins (T. Hunter, Cell 1987 50:823-829). Specifically,tyrosine kinases phosphorylate proteins on the phenolic moiety oftyrosine residues. The tyrosine kinase family includes members thatcontrol cell growth, migration, and differentiation. Abnormal kinaseactivity has been implicated in a variety of human diseases includingcancers, autoimmune and inflammatory diseases. Since protein kinases areamong the key regulators of cell signaling they provide a target tomodulate cellular function with small molecular kinase inhibitors andthus make good drug design targets. In addition to treatment ofkinase-mediated disease processes, selective and efficacious inhibitorsof kinase activity are also useful for investigation of cell signalingprocesses and identification of other cellular targets of therapeuticinterest.

There is good evidence that B-cells play a key role in the pathogenesisof autoimmune and/or inflammatory disease. Protein-based therapeuticsthat deplete B cells such as Rituxan are effective againstautoantibody-driven inflammatory diseases such as rheumatoid arthritis(Rastetter et al. Annu Rev Med 2004 55:477). Therefore inhibitors of theprotein kinases that play a role in B-cell activation should be usefultherapeutics for B-cell mediated disease pathology such as autoantibodyproduction.

Signaling through the B-cell receptor (BCR) controls a range of B-cellresponses including proliferation and differentiation into matureantibody producing cells. The BCR is a key regulatory point for B-cellactivity and aberrant signaling can cause deregulated B-cellproliferation and formation of pathogenic autoantibodies that lead tomultiple autoimmune and/or inflammatory diseases. Bruton's TyrosineKinase (Btk) is a non-BCR associated kinase that is membrane proximaland immediately downstream from BCR. Lack of Btk has been shown to blockBCR signaling and therefore inhibition of Btk could be a usefultherapeutic approach to block B-cell mediated disease processes.

Btk is a member of the Tec family of tyrosine kinases, and has beenshown to be a critical regulator of early B-cell development and matureB-cell activation and survival (Khan et al. Immunity 1995 3:283;Ellmeier et al. J. Exp. Med. 2000 192:1611). Mutation of Btk is humansleads to the condition X-linked agammaglobulinemia (XLA) (reviewed inRosen et al. New Eng. J. Med. 1995 333:431 and Lindvall et al. Immunol.Rev. 2005 203:200). These patients are immunocompromised and showimpaired maturation of B-cells, decreased immunoglobulin and peripheralB-cell levels, diminished T-cell independent immune responses as well asattenuated calcium mobilization following BCR stimulation.

Evidence for a role for Btk in autoimmune and inflammatory diseases hasalso been provided by Btk-deficient mouse models. In preclinical murinemodels of systemic lupus erythematosus (SLE), Btk-deficient mice showmarked amelioration of disease progression. In addition, Btk-deficientmice are resistant to collagen-induced arthritis (Jansson and HolmdahlClin. Exp. Immunol. 1993 94:459). A selective Btk inhibitor has beendemonstrated dose-dependent efficacy in a mouse arthritis model (Z. Panet al., Chem. Med Chem. 2007 2:58-61).

Btk is also expressed by cells other than B-cells that may be involvedin disease processes. For example, Btk is expressed by mast cells andBtk-deficient bone marrow derived mast cells demonstrate impairedantigen induced degranulation (Iwaki et al. J. Biol. Chem. 2005280:40261). This shows Btk could be useful to treat pathological mastcells responses such as allergy and asthma. Also monocytes from XLApatients, in which Btk activity is absent, show decreased TNF alphaproduction following stimulation (Horwood et al. J Exp Med 197:1603,2003). Therefore TNF alpha mediated inflammation could be modulated bysmall molecular Btk inhibitors. Also, Btk has been reported to play arole in apoptosis (Islam and Smith Immunol. Rev. 2000 178:49) and thusBtk inhibitors would be useful for the treatment of certain B-celllymphomas and leukemias (Feldhahn et al. J. Exp. Med. 2005 201:1837).

SUMMARY OF THE INVENTION

The present application provides the Btk inhibitor compounds of FormulaI, methods of use thereof, as described herein below:

The application provides a compound of Formula I,

wherein:

R¹ is H or halo;

R² is H, halo, or cyano;

R³ is R⁴ or R⁵;

-   -   R⁴ is halo or cyano;    -   R⁵ is phenyl, heteroaryl, —C(═O)R^(5′), lower alkyl, or benzyl,        optionally substituted with one or more R^(5′);    -   R^(5′) is lower alkyl, cyano, hydroxyl, heterocycloalkyl,        phenyl, amino, alkyl amino, dialkyl amino, or lower alkoxy; and

X is lower alkyl or halo;

or a pharmaceutically acceptable salt thereof.

The application provides a method for treating an inflammatory and/orautoimmune condition comprising administering to a patient in needthereof a therapeutically effective amount of the compound of Formula I.

The application provides a pharmaceutical composition comprising thecompound of Formula I, admixed with at least one pharmaceuticallyacceptable carrier, excipient or diluent.

DETAILED DESCRIPTION OF THE INVENTION Definitions

The phrase “a” or “an” entity as used herein refers to one or more ofthat entity; for example, a compound refers to one or more compounds orat least one compound. As such, the terms “a” (or “an”), “one or more”,and “at least one” can be used interchangeably herein.

The phrase “as defined herein above” refers to the broadest definitionfor each group as provided in the Summary of the Invention or thebroadest claim. In all other embodiments provided below, substituentswhich can be present in each embodiment and which are not explicitlydefined retain the broadest definition provided in the Summary of theInvention.

As used in this specification, whether in a transitional phrase or inthe body of the claim, the terms “comprise(s)” and “comprising” are tobe interpreted as having an open-ended meaning. That is, the terms areto be interpreted synonymously with the phrases “having at least” or“including at least”. When used in the context of a process, the term“comprising” means that the process includes at least the recited steps,but may include additional steps. When used in the context of a compoundor composition, the term “comprising” means that the compound orcomposition includes at least the recited features or components, butmay also include additional features or components.

As used herein, unless specifically indicated otherwise, the word “or”is used in the “inclusive” sense of “and/or” and not the “exclusive”sense of “either/or”.

The term “independently” is used herein to indicate that a variable isapplied in any one instance without regard to the presence or absence ofa variable having that same or a different definition within the samecompound. Thus, in a compound in which R″ appears twice and is definedas “independently carbon or nitrogen”, both R″s can be carbon, both R″scan be nitrogen, or one R″ can be carbon and the other nitrogen.

When any variable occurs more than one time in any moiety or formuladepicting and describing compounds employed or claimed in the presentinvention, its definition on each occurrence is independent of itsdefinition at every other occurrence. Also, combinations of substituentsand/or variables are permissible only if such compounds result in stablecompounds.

The symbols “*” at the end of a bond or “—” drawn through a bond eachrefer to the point of attachment of a functional group or other chemicalmoiety to the rest of the molecule of which it is a part. Thus, forexample:

A bond drawn into ring system (as opposed to connected at a distinctvertex) indicates that the bond may be attached to any of the suitablering atoms.

The term “optional” or “optionally” as used herein means that asubsequently described event or circumstance may, but need not, occur,and that the description includes instances where the event orcircumstance occurs and instances in which it does not. For example,“optionally substituted” means that the optionally substituted moietymay incorporate a hydrogen atom or a substituent.

The phrase “optional bond” means that the bond may or may not bepresent, and that the description includes single, double, or triplebonds. If a substituent is designated to be a “bond” or “absent”, theatoms linked to the substituents are then directly connected.

The term “about” is used herein to mean approximately, in the region of,roughly, or around. When the term “about” is used in conjunction with anumerical range, it modifies that range by extending the boundariesabove and below the numerical values set forth. In general, the term“about” is used herein to modify a numerical value above and below thestated value by a variance of 20%.

Certain compounds of Formulae I may exhibit tautomerism. Tautomericcompounds can exist as two or more interconvertable species. Prototropictautomers result from the migration of a covalently bonded hydrogen atombetween two atoms. Tautomers generally exist in equilibrium and attemptsto isolate an individual tautomers usually produce a mixture whosechemical and physical properties are consistent with a mixture ofcompounds. The position of the equilibrium is dependent on chemicalfeatures within the molecule. For example, in many aliphatic aldehydesand ketones, such as acetaldehyde, the keto form predominates while, inphenols, the enol form predominates. Common prototropic tautomersinclude keto/enol (—C(═O)—CH— ⇄—C(—OH)═CH—), amide/imidic acid(—C(═O)—NH— ⇄—C(—OH)═N—) and amidine (—C(═NR)—NH— ⇄—C(—NHR)═N—)tautomers. The latter two are particularly common in heteroaryl andheterocyclic rings and the present invention encompasses all tautomericforms of the compounds.

Technical and scientific terms used herein have the meaning commonlyunderstood by one of skill in the art to which the present inventionpertains, unless otherwise defined. Reference is made herein to variousmethodologies and materials known to those of skill in the art. Standardreference works setting forth the general principles of pharmacologyinclude Goodman and Gilman's The Pharmacological Basis of Therapeutics,10^(th) Ed., McGraw Hill Companies Inc., New York (2001). Any suitablematerials and/or methods known to those of skill can be utilized incarrying out the present invention. However, preferred materials andmethods are described. Materials, reagents and the like to whichreference are made in the following description and examples areobtainable from commercial sources, unless otherwise noted.

The definitions described herein may be appended to formchemically-relevant combinations, such as “heteroalkylaryl”,“haloalkylheteroaryl”, “arylalkylheterocyclyl”, “alkylcarbonyl”,“alkoxyalkyl”, and the like. When the term “alkyl” is used as a suffixfollowing another term, as in “phenylalkyl”, or “hydroxyalkyl”, this isintended to refer to an alkyl group, as defined above, being substitutedwith one to two substituents selected from the other specifically-namedgroup. Thus, for example, “phenylalkyl” refers to an alkyl group havingone to two phenyl substituents, and thus includes benzyl, phenylethyl,and biphenyl. An “alkylaminoalkyl” is an alkyl group having one to twoalkylamino substituents, “Hydroxyalkyl” includes 2-hydroxyethyl,2-hydroxypropyl, 1-(hydroxymethyl)-2-methylpropyl, 2-hydroxybutyl,2-dihydroxybutyl, 2-(hydroxymethyl), 3-hydroxypropyl, and so forth.Accordingly, as used herein, the term “hydroxyalkyl” is used to define asubset of heteroalkyl groups defined below. The term -(ar)alkyl refersto either an unsubstituted alkyl or an aralkyl group. The term(hetero)aryl or (het)aryl refers to either an aryl or a heteroarylgroup.

The term “spirocycloalkyl”, as used herein, means a spirocycliccycloalkyl group, such as, for example, spiro[3.3]heptane. The termspiroheterocycloalkyl, as used herein, means a spirocyclicheterocycloalkyl, such as, for example, 2,6-diazaspiro[3.3]heptane.

The term “acyl” as used herein denotes a group of formula —C(═O)Rwherein R is hydrogen or lower alkyl as defined herein. The term or“alkylcarbonyl” as used herein denotes a group of formula C(═O)R whereinR is alkyl as defined herein. The term C₁₋₆ acyl refers to a group—C(═O)R contain 6 carbon atoms. The term “arylcarbonyl” as used hereinmeans a group of formula C(═O)R wherein R is an aryl group; the term“benzoyl” as used herein an “arylcarbonyl” group wherein R is phenyl.

The term “ester” as used herein denotes a group of formula —C(═O)ORwherein R is lower alkyl as defined herein.

The term “alkyl” as used herein denotes an unbranched or branched chain,saturated, monovalent hydrocarbon residue containing 1 to 10 carbonatoms. The term “lower alkyl” denotes a straight or branched chainhydrocarbon residue containing 1 to 6 carbon atoms. “C₁₋₁₀ alkyl” asused herein refers to an alkyl composed of 1 to 10 carbons. Examples ofalkyl groups include, but are not limited to, lower alkyl groups includemethyl, ethyl, propyl, i-propyl, n-butyl, i-butyl, t-butyl or pentyl,isopentyl, neopentyl, hexyl, heptyl, and octyl.

When the term “alkyl” is used as a suffix following another term, as in“phenylalkyl,” or “hydroxyalkyl,” this is intended to refer to an alkylgroup, as defined above, being substituted with one to two substituentsselected from the other specifically-named group. Thus, for example,“phenylalkyl” denotes the radical R′R″—, wherein R′ is a phenyl radical,and R″ is an alkylene radical as defined herein with the understandingthat the attachment point of the phenylalkyl moiety will be on thealkylene radical. Examples of arylalkyl radicals include, but are notlimited to, benzyl, phenylethyl, 3-phenylpropyl. The terms “arylalkyl”or “aralkyl” are interpreted similarly except R′ is an aryl radical. Theterms “(het)arylalkyl” or “(het)aralkyl” are interpreted similarlyexcept R′ is optionally an aryl or a heteroaryl radical.

The terms “haloalkyl” or “halo-lower alkyl” or “lower haloalkyl” refersto a straight or branched chain hydrocarbon residue containing 1 to 6carbon atoms wherein one or more carbon atoms are substituted with oneor more halogen atoms.

The term “alkylene” or “alkylenyl” as used herein denotes a divalentsaturated linear hydrocarbon radical of 1 to 10 carbon atoms (e.g.,(CH₂)_(n)) or a branched saturated divalent hydrocarbon radical of 2 to10 carbon atoms (e.g., —CHMe- or —CH₂CH(i-Pr)CH₂—), unless otherwiseindicated. Except in the case of methylene, the open valences of analkylene group are not attached to the same atom. Examples of alkyleneradicals include, but are not limited to, methylene, ethylene,propylene, 2-methyl-propylene, 1,1-dimethyl-ethylene, butylene,2-ethylbutylene.

The term “alkoxy” as used herein means an —O-alkyl group, wherein alkylis as defined above such as methoxy, ethoxy, n-propyloxy, i-propyloxy,n-butyloxy, i-butyloxy, t-butyloxy, pentyloxy, hexyloxy, including theirisomers. “Lower alkoxy” as used herein denotes an alkoxy group with a“lower alkyl” group as previously defined. “C₁₋₁₀ alkoxy” as used hereinrefers to an-O-alkyl wherein alkyl is C₁₋₁₀.

The term “PCy₃” refers to a phosphine t substituted with three cyclicmoieties.

The terms “haloalkoxy” or “halo-lower alkoxy” or “lower haloalkoxy”refers to a lower alkoxy group, wherein one or more carbon atoms aresubstituted with one or more halogen atoms.

The term “hydroxyalkyl” as used herein denotes an alkyl radical asherein defined wherein one to three hydrogen atoms on different carbonatoms is/are replaced by hydroxyl groups.

The terms “alkylsulfonyl” and “arylsulfonyl” as used herein refers to agroup of formula —S(═O)₂R wherein R is alkyl or aryl respectively andalkyl and aryl are as defined herein. The term “heteroalkylsulfonyl” asused herein refers herein denotes a group of formula —S(═O)₂R wherein Ris “heteroalkyl” as defined herein.

The terms “alkylsulfonylamino” and “arylsulfonylamino” as used hereinrefers to a group of formula —NR′S(═O)₂R wherein R is alkyl or arylrespectively, R′ is hydrogen or C₁₋₃ alkyl, and alkyl and aryl are asdefined herein.

The term “cycloalkyl” as used herein refers to a saturated carbocyclicring containing 3 to 8 carbon atoms, i.e. cyclopropyl, cyclobutyl,cyclopentyl, cyclohexyl, cycloheptyl or cyclooctyl. “C₃₋₇cycloalkyl” asused herein refers to a cycloalkyl composed of 3 to 7 carbons in thecarbocyclic ring.

The term carboxy-alkyl as used herein refers to an alkyl moiety whereinone, hydrogen atom has been replaced with a carboxyl with theunderstanding that the point of attachment of the heteroalkyl radical isthrough a carbon atom. The term “carboxy” or “caboxyl” refers to a —CO₂Hmoiety.

The term “heteroaryl” or “heteroaromatic” as used herein means amonocyclic or bicyclic radical of 5 to 12 ring atoms having at least onearomatic or partially unsaturated ring containing four to eight atomsper ring, incorporating one or more N, O, or S heteroatoms, theremaining ring atoms being carbon, with the understanding that theattachment point of the heteroaryl radical will be on an aromatic orpartially unsaturated ring. As well known to those skilled in the art,heteroaryl rings have less aromatic character than their all-carboncounter parts. Thus, for the purposes of the invention, a heteroarylgroup need only have some degree of aromatic character. Examples ofheteroaryl moieties include monocyclic aromatic heterocycles having 5 to6 ring atoms and 1 to 3 heteroatoms include, but is not limited to,pyridinyl, pyrimidinyl, pyrazinyl, oxazinyl, pyrrolyl, pyrazolyl,imidazolyl, oxazolyl, 4,5-Dihydro-oxazolyl,5,6-Dihydro-4H-[1,3]oxazolyl, isoxazole, thiazole, isothiazole,triazoline, thiadiazole and oxadiaxoline which can optionally besubstituted with one or more, preferably one or two substituentsselected from hydroxy, cyano, alkyl, alkoxy, thio, lower haloalkoxy,alkylthio, halo, lower haloalkyl, alkylsulfinyl, alkylsulfonyl, halogen,amino, alkylamino, dialkylamino, aminoalkyl, alkylaminoalkyl, anddialkylaminoalkyl, nitro, alkoxycarbonyl and carbamoyl, alkylcarbamoyl,dialkylcarbamoyl, arylcarbamoyl, alkylcarbonylamino andarylcarbonylamino. Examples of bicyclic moieties include, but are notlimited to, quinolinyl, isoquinolinyl, benzofuryl, benzothiophenyl,benzoxazole, benzisoxazole, benzothiazole, naphthyridinyl,5,6,7,8-Tetrahydro-[1,6]naphthridinyl, and benzisothiazole. Bicyclicmoieties can be optionally substituted on either ring, however the pointof attachment is on a ring containing a heteroatom.

The term “heterocyclyl”, “heterocycloalkyl” or “heterocycle” as usedherein denotes a monovalent saturated cyclic radical, consisting of oneor more rings, preferably one to two rings, including spirocyclic ringsystems, of three to eight atoms per ring, incorporating one or morering heteroatoms (chosen from N, O or S(O)₀₋₂), and which can optionallybe independently substituted with one or more, preferably one or twosubstituents selected from hydroxy, oxo, cyano, lower alkyl, loweralkoxy, lower haloalkoxy, alkylthio, halo, lower haloalkyl,hydroxyalkyl, nitro, alkoxycarbonyl, amino, alkylamino, alkylsulfonyl,arylsulfonyl, alkylaminosulfonyl, arylaminosulfonyl, alkylsulfonylamino,arylsulfonylamino, alkylaminocarbonyl, arylaminocarbonyl,alkylcarbonylamino, arylcarbonylamino, and ionic forms thereof, unlessotherwise indicated. Examples of heterocyclic radicals include, but arenot limited to, morpholinyl, piperazinyl, piperidinyl, azetidinyl,pyrrolidinyl, hexahydroazepinyl, oxetanyl, tetrahydrofuranyl,tetrahydrothiophenyl, oxazolidinyl, thiazolidinyl, isoxazolidinyl,tetrahydropyranyl, thiomorpholinyl, quinuclidinyl and imidazolinyl, andionic forms thereof. Examples may also be bicyclic, such as, forexample, 3,8-diaza-bicyclo[3.2.1]octane, 2,5-diaza-bicyclo[2.2.2]octane,or octahydro-pyrazino[2,1-c][1,4]oxazine.

Inhibitors of Btk

The application provides a compound of Formula I,

wherein:

R¹ is H or halo;

R² is H, halo, or cyano;

R³ is R⁴ or R⁵;

-   -   R⁴ is halo or cyano;    -   R⁵ is phenyl, heteroaryl, —C(═O)R^(5′), lower alkyl, or benzyl,        optionally substituted with one or more R^(5′);    -   R^(5′) is lower alkyl, cyano, hydroxyl, heterocycloalkyl,        phenyl, amino, alkyl amino, dialkyl amino, or lower alkoxy; and

X is lower alkyl or halo;

or a pharmaceutically acceptable salt thereof.

The application provides a compound of Formula I, wherein X is methyl.

The application provides a compound of Formula I, wherein X is halo.

The application provides a compound of Formula I, wherein X is methyland R⁵ is heteroaryl, optionally substituted with one or more R^(5′).

The application provides a compound of Formula I, wherein R⁵ isthiophenyl, optionally substituted with one or more R^(5′).

The application provides a compound of Formula I, wherein R⁵ is pyridyl,optionally substituted with one or more R^(5′).

The application provides any of the above compounds of Formula I,wherein R¹ is F and R² is F.

-   -   The application alternatively provides any of the above        compounds of Formula I, wherein R¹ is H and R² is cyano.    -   The application provides a compound of Formula I, wherein R⁵ is        —C(═O)R^(5′).    -   The application provides a compound of Formula I, wherein R^(5′)        is morpholinyl, piperidinyl, loweralyl piperidinyl, or lower        alkoxy.    -   The application provides a compound of Formula I, wherein R¹ is        F and R² is F.    -   The application provides a compound of Formula I, wherein R¹ is        H and R² is cyano.    -   The application provides a compound of Formula I, wherein R⁵ is        phenyl or benzyl, optionally substituted with one or more        R^(5′).    -   The application provides a compound of Formula I, wherein R⁵ is        lower alkylene, optionally substituted with one or more R^(5′).    -   The application provides either of the above compounds of        Formula I, wherein R¹ is F and R² is F.

The application alternatively provides either of the above compounds ofFormula I, wherein R¹ is H and R² is cyano.

The application provides a compound of Formula I, selected from thegroup consisting of:

-   {5-Amino-1-[4-(2,3-difluoro-phenoxy)-2-methyl-phenyl]-1H-pyrazol-4-yl}-(4-bromo-1H-indol-2-yl)-methanone;-   {5-Amino-1-[4-(2,3-difluoro-phenoxy)-2-methyl-phenyl]-1H-pyrazol-4-yl}-(4-phenyl-1H-indol-2-yl)-methanone;-   {5-Amino-1-[4-(2,3-difluoro-phenoxy)-2-methyl-phenyl]-1H-pyrazol-4-yl}-(4-thiophen-3-yl-1H-indol-2-yl)-methanone;-   {5-Amino-1-[4-(2,3-difluoro-phenoxy)-2-methyl-phenyl]-1H-pyrazol-4-yl}-(4-pyrazol-1-yl-1H-indol-2-yl)-methanone;-   {5-Amino-1-[4-(2,3-difluoro-phenoxy)-2-methyl-phenyl]-1H-pyrazol-4-yl}-(4-thiophen-2-yl-1H-indol-2-yl)-methanone;-   {5-Amino-1-[4-(2,3-difluoro-phenoxy)-2-methyl-phenyl]-1H-pyrazol-4-yl}-[4-(morpholine-4-carbonyl)-1H-indol-2-yl]-methanone;-   {5-Amino-1-[4-(2,3-difluoro-phenoxy)-2-methyl-phenyl]-1H-pyrazol-4-yl}-[4-(1-methyl-1H-pyrazol-4-yl)-1H-indol-2-yl]-methanone;-   {5-Amino-1-[4-(2,3-difluoro-phenoxy)-2-methyl-phenyl]-1H-pyrazol-4-yl}-(4-pyridin-2-yl-1H-indol-2-yl)-methanone;-   {5-Amino-1-[4-(2,3-difluoro-phenoxy)-2-methyl-phenyl]-1H-pyrazol-4-yl}-(4-benzyl-1H-indol-2-yl)-methanone;-   {5-Amino-1-[4-(2,3-difluoro-phenoxy)-2-methyl-phenyl]-1H-pyrazol-4-yl}-[4-(1H-pyrazol-4-yl)-1H-indol-2-yl]-methanone;-   3-(2-{5-Amino-1-[4-(2,3-difluoro-phenoxy)-2-methyl-phenyl]-1H-pyrazole-4-carbonyl}-1H-indol-4-yl)-benzonitrile;-   {5-Amino-1-[4-(2,3-difluoro-phenoxy)-2-methyl-phenyl]-1H-pyrazol-4-yl}-[4-(3-chloro-phenyl)-1H-indol-2-yl]-methanone;-   3-(2-{5-Amino-1-[4-(2,3-difluoro-phenoxy)-2-methyl-phenyl]-1H-pyrazole-4-carbonyl}-1H-indol-4-ylmethyl)-benzonitrile;-   3-(4-{5-Amino-4-[4-(1H-pyrazol-4-yl)-1H-2-carbonyl]-pyrazol-1-yl}-3-methyl-phenoxy)-benzonitrile;-   3-(4-{5-Amino-4-[4-(morpholine-4-carbonyl)-1H-indole-2-carbonyl]-pyrazol-1-yl}-3-methyl-phenoxy)-benzonitrile;-   3-(4-{5-Amino-4-[4-(4-methyl-piperazine-1-carbonyl)-1H-indole-2-carbonyl]-pyrazol-1-yl}-3-methyl-phenoxy)-benzonitrile;-   3-(4-{5-Amino-4-[4-(3-methoxy-benzyl)-1H-indole-2-carbonyl]-pyrazol-1-yl}-3-methyl-phenoxy)-benzonitrile;-   2-{5-Amino-1-[4-(2,3-difluoro-phenoxy)-2-methyl-phenyl]-1H-pyrazole-4-carbonyl}-1H-indole-4-carboxylic    acid methyl ester;-   {5-Amino-1-[4-(2,3-difluoro-phenoxy)-2-methyl-phenyl]-1H-pyrazol-4-yl}-(4-morpholin-4-ylmethyl-1H-indol-2-yl)-methanone;-   3-{4-[5-Amino-4-(4-cyanomethyl-1H-indole-2-carbonyl)-pyrazol-1-yl]-3-methyl-phenoxy}-benzonitrile;    and-   2-{5-Amino-1-[4-(3-cyano-phenoxy)-2-methyl-phenyl]-1H-pyrazole-4-carbonyl}-1H-indole-4-carboxylic    acid methylamide.    -   The application provides a method for treating an inflammatory        and/or autoimmune condition comprising administering to a        patient in need thereof a therapeutically effective amount of        the compound of Formula I.    -   The application provides a method for treating rheumatoid        arthritis comprising administering to a patient in need thereof        a therapeutically effective amount of the compound of Formula I.    -   The application provides a method for treating asthma comprising        administering to a patient in need thereof a therapeutically        effective amount of the compound of Formula I.    -   The application provides a method for treating cancer comprising        administering to a patient in need thereof a therapeutically        effective amount of the compound of Formula I.    -   The application provides a pharmaceutical composition comprising        the compound of Formula I.    -   The application provides a pharmaceutical composition comprising        the compound of Formula I, admixed with at least one        pharmaceutically acceptable carrier, excipient car diluent.    -   The application provides the use of the compound of formula I as        therapeutically active substance.    -   The application provides a use of the compound of formula I in        the manufacture of a medicament for the treatment of an        inflammatory disorder.    -   The application provides a use of the compound of formula I in        the manufacture of a medicament for the treatment of an        autoimmune disorder.    -   The application provides a use of the compound of formula I in        the manufacture of a medicament for the treatment of rheumatoid        arthritis.    -   The application provides a use of the compound of formula I in        the manufacture of a medicament for the treatment of asthma.    -   The application provides the use of a compound as described        above for the treatment of inflammatory and/or autoimmune        condition.

The application provides the use of a compound as described above forthe treatment of rheumatoid arthritis.

The application provides the use of a compound as described above forthe treatment of asthma.

The application provides the use of a compound as described above forthe treatment of inflammatory and/or autoimmune condition.

The application provides the use of a compound as described above forthe treatment of rheumatoid arthritis.

The application provides the use of a compound as described above forthe treatment of asthma.

The application provides a compound as described above for use in thetreatment of inflammatory and/or autoimmune condition.

The application provides a compound as described above for use in thetreatment of rheumatoid arthritis.

The application provides a compound as described above for use in thetreatment of asthma.

The application provides a compound, method, or composition as describedherein.

Compounds and Preparation

Examples of representative compounds encompassed by the presentinvention and within the scope of the invention are provided in thefollowing Table. These examples and preparations which follow areprovided to enable those skilled in the art to more clearly understandand to practice the present invention. They should not be considered aslimiting the scope of the invention, but merely as being illustrativeand representative thereof.

In general, the nomenclature used in this Application is based onAUTONOM™ v.4.0, a Beilstein Institute computerized system for thegeneration of IUPAC systematic nomenclature. If there is a discrepancybetween a depicted structure and a name given that structure, thedepicted structure is to be accorded more weight. In addition, if thestereochemistry of a structure or a portion of a structure is notindicated with, for example, bold or dashed lines, the structure orportion of the structure is to be interpreted as encompassing allstereoisomers of it.

TABLE I depicts examples of compounds according to genetic Formula I:

TABLE I Compound Nomenclature Structure 1 {5-Amino-1- [4-(2,3- difluoro-phenoxy)-2- methyl- phenyl]-1H- pyrazol-4-yl}- (4-bromo-1H- indol-2-yl)-methanone

2 {5-Amino-1- [4-(2,3- difluoro- phenoxy)-2- methyl- phenyl]-1H-pyrazol-4-yl}- (4-phenyl-1H- indol-2-yl)- methanone

3 {5-Amino-1- [4-(2,3- difluoro- phenoxy)-2- methyl- phenyl]-1H-pyrazol-4-yl}- (4-thiophen-3- yl-1H-indol-2- yl)-methanone

4 {5-Amino-1- [4-(2,3- difluoro- phenoxy)-2- methyl- phenyl]-1H-pyrazol-4-yl}- (4-pyrazol-1- yl-1H-indol-2- yl)-methanone

5 {5-Amino-1- [4-(2,3- difluoro- phenoxy)-2- methyl- phenyl]-1H-pyrazol-4-yl}- (4-thiophen-2- yl-1H-indol-2- yl)-methanone

6 {5-Amino-1- [4-(2,3- difluoro- phenoxy)-2- methyl- phenyl]-1H-pyrazol-4-yl}- [4- (morpholine-4- carbonyl)-1H- indol-2-yl]- methanone

7 {5-Amino-1- [4-(2,3- difluoro- phenoxy)-2- methyl- phenyl]-1H-pyrazol-4-yl}- [4-(1-methyl- 1H-pyrazol-4- yl)-1H- indol-2-yl]-methanone

8 {5-Amino-1- [4-(2,3- difluoro- phenoxy)-2- methyl- phenyl]-1H-pyrazol-4-yl}- (4-pyridin-2- yl-1H-indol-2- yl)-methanone

9 {5-Amino-1- [4-(2,3- difluoro- phenoxy)-2- methyl- phenyl]-1H-pyrazol-4-yl}- (4-benzyl-1H- indol-2-yl)- methanone

10 {5-Amino-1- [4-(2,3- difluoro- phenoxy)-2- methyl- phenyl]-1H-pyrazol-4-yl}- [4-(1H- pyrazol-4-yl)- 1H-indol-2- yl]-methanone

11 3-(2-{5- Amino-1-[4- (2,3-difluoro- phenoxy)-2- methyl- phenyl]-1H-pyrazole-4- carbonyl}-1H- indol-4-yl)- benzonitrile

12 {5-Amino-1- [4-(2,3- difluoro- phenoxy)-2- methyl- phenyl]-1H-pyrazol-4-yl}- [4-(3-chloro- phenyl)-1H- indol-2-yl]- methanone

13 3-(2-{5- Amino-1-[4- (2,3-difluoro- phenoxy)-2- methyl- phenyl]-1H-pyrazole-4- carbonyl}-1H- indol-4- ylmethyl)- benzonitrile

14 3-(4-{5- Amino-4-[4- (1H-pyrazol-4- yl)-1H-indole- 2-carbonyl]-pyrazol-1-yl}- 3-methyl- phenoxy)- benzonitrile

15 3-(4-{5- Amino-4-[4- (morpholine-4- carbonyl)-1H- indole-2-carbonyl]- pyrazol-1-yl}- 3-methyl- phenoxy)- benzonitrile

16 3-(4-{5- Amino-4-[4- (4-methyl- piperazine-1- carbonyl)-1H- indole-2-carbonyl]- pyrazol-1-yl}- 3-methyl- phenoxy)- benzonitrile

17 3-(4-{5- Amino-4-[4- (3-methoxy- benzyl)-1H- indole-2- carbonyl]-pyrazol-1-yl}- 3-methyl- phenoxy)- benzonitrile

18 2-{5-Amino- 1-[4-(2,3- difluoro- phenoxy)-2- methyl- phenyl]-1H-pyrazole-4- carbonyl}-1H- indole-4- carboxylic acid methyl ester

19 {5-Amino-1- [4-(2,3- difluoro- phenoxy)-2- methyl- phenyl]-1H-pyrazol-4-yl}- (4-morpholin- 4-ylmethyl- 1H-indol-2- yl)-methanone

20 3-{4-[5- Amino-4-(4- cyanomethyl- 1H-indole-2- carbonyl)-pyrazol-1-yl]- 3-methyl- phenoxy}- benzonitrile

21 2-{5-Amino- 1-[4-(3-cyano- phenoxy)-2- methyl- phenyl]-1H-pyrazole-4- carbonyl}-1H- indole-4- carboxylic acid methylamide

General Synthetic Schemes

The compounds of the present invention may be prepared by anyconventional means. Suitable processes for synthesizing these compoundsare provided in the examples. Generally, compounds of the invention maybe prepared according to the schemes below.

Compounds of formula 11, where R1 and R2 are as described above in thegenus of formula 1, may be prepared using the route outlined inScheme 1. According to this procedure, the compound of formula 1,4-bromoindole, which is commercially available, may be converted to thephenylsulfonamide of formula 2. Treatment with strong base and carbondioxide provide carboxylic acid 3 which is converted to the methyl ester5 via the acid chloride 4. The ester 5 may then be reacted with an anionderived from acetonitrile to give the cyanoacetyl derivative of formula6. Reaction with dimethylformamide dimethyl acetal provides theacrylonitrile derivative 7 and this reacts with the phenyl hydrazinederivative of formula 8 to give the aminopyrazole of formula 9. The R2group may then be installed using a transition-metal catalyzed couplingreaction or a nucleophilic aromatic substitution reaction as outlinedbelow, to give the compound of formula 10. Removal of the phenylsulfonylprotective group then provides the compound of the invention of formula11.

4-Bromoindole, the compound of formula 1, may be conveniently treatedwith a base such as sodium hydride in an inert solvent such astetrahydrofuran at a temperature around 0° C. to generate thecorresponding anion. This may be treated with benzenesulfonyl chlorideand the mixture stirred at room temperature for about an hour to givethe benzenesulfonamide derivative of formula 2.

The compound of formula 2 may then be treated with n-butyl-lithium intetrahydrofuran at low temperature, such as at about −78° C., and thecorresponding anion treated with excess solid carbon dioxide to give thecarboxylic acid of formula 3.

The conversion of the carboxylic acid of formula 3 to the methyl esterof formula 5 may be effected using one of a variety of methods that arewell known to one of average skill in the art of organic synthesis. Manysuitable approaches are enumerated in Greene's Protective Groups inOrganic Synthesis [Wuts, P. G. M and Greene, T. W., 4th Edition,Wiley-Interscience, New York, 2006, pages 553 et seq.] For example, thetransformation may be conveniently carried out by treating thecarboxylic acid of formula 3 with a chlorinating agent such as thionylchloride either neat or in an inert solvent such as benzene at atemperature between about 50° C. and about the reflux temperature. Theresulting acid chloride of formula 4 may then be treated with methanolin the presence of a base such as triethylamine or diisopropylethylamineor pyridine either using methanol as solvent or in an inert solvent suchas tetrahydrofuran at about room temperature.

Specific conditions for the preparation of the compound of formula 5 maybe found in the literature, in Mahboobi, S. et al. J. Med. Chem. 2006,49, 3101-3115.

The compound of formula 5 may be conveniently converted to thecyanoacetyl derivative of formula 6 by treating it with a mixture ofacetonitrile and a strong base such as lithium diisopropylamide orlithium hexamethyldisilazide in a solvent such as tetrahydrofuran at lowtemperature, such as at about −78° C. Conditions for such a reaction maybe found in the patent literature, for example in Taka, N. et al. US20120208811 Page 163.

The compound of formula 6 may be converted to the acrylonitrilederivative of formula 7 by treatment with N,N-dimethylformamide dimethylacetal in an inert solvent such as an aromatic hydrocarbon (e.g.,toluene) or tetrahydrofuran at about room temperature. Conditions forsuch a reaction may be found in the patent literature, for example inTaka, N. et al. US 20120208811 page 132.

The acrylonitrile derivative of formula 7 may be converted to theaminopyrazole derivative of formula 9 by treatment with an intermediateof formula 8, where R1 is as described above in the genus of formula I,in an alcoholic solvent such as methanol or ethanol or isopropanol, atabout the reflux temperature of the solvent. Conditions for such areaction may be found in the patent literature, for example in Taka, N.et al. US 20120208811 Page 94.

The reaction of a compound of formula 9 with a compound of formula R₂—X,where X represents boronic acid, boronate ester, potassiumtrifluoroborate, trimethyltin car tri-n-butyl-tin, to give a compound offormula 10 can be effected using Suzuki or Stile or Negishi couplingconditions which are well known to one of average skill in the art. Forexample, in a Suzuki reaction, the reaction can be conveniently carriedout by reacting a compound of formula 9 with a compound of formulaR₂—B(OH)₂, in a convenient inert solvent such as a polar aprotic solvent(e.g., N,N-dimethylformamide) or an ether (e.g., dioxane) or water, orindeed in a mixture of such solvents, in the presence of a catalyticamount of a palladium(0) precursor (for example, palladium(II) acetateor bis(triphenylphosphine)palladium(II) chloride), in the optionaladditional presence of a catalytic amount of a phosphine ligand, forexample tri-o-tolylphosphine or tri-tert-butylphosphine, oralternatively in the presence of a preformed complex of palladium(0)with a phosphine ligand such as bis(tri-cyclohexylphosphine)palladium,tetrakis(triphenylphosphine) palladium(0) or[1,1-bis(diphenylphosphino)ferrocene]dichloropalladium(II)), and also inthe presence of an inorganic base, for example, an alkali metalcarbonate, bicarbonate or phosphate (e.g., potassium phosphate or sodiumcarbonate) at a temperature between about room temperature and about 100degrees, and preferably between about room temperature and about 50degrees. The Suzuki reaction is familiar to one of ordinary skill in theart of organic synthesis, and has been reviewed several times, notablyin Miyaura, N.; Suzuki, A. Chem. Rev. 1995, 95, 2457-2483 and, morerecently, in Alonso, F.; Beletskaya, I. P.; Yus, M. Tetrahedron 2008,64, 3047-3101. Examples of specific conditions useful for Suzukicoupling may be found in many references in the literature including:Tiede, S. et al. Angew. Chem. Intl. Edn. 2010, 49, 3972-3975; Schmidt,A. and Rahirni, A. Chem. Commun. 2010, 46, 2995-2997; Lee, S. H. et al.US 20100063281; and Tobisu, M. et al, J. Org. Chem. 2010, 75, 4835-4840(Supporting Information). Stile coupling is well known to one of averageskill in the field of organic synthesis, and may be used as analternative to the Suzuki coupling, examples of conditions for whichhave been provided above. Stile coupling has been reviewed, including inFarina, V. et al. Org. Reactions 1997, 50, 1-652. Examples of specificconditions that have been used for Stille coupling may be found in theliterature, for example in Littke, A, F. et al. J. Am. Chem. Soc. 2002,124, 5343-6348; in Alberati-Giani, D. et al. U.S. Pat. No. 7,462,617;and in Robl, J. A. U.S. Pat. No. 5,072,023. For example, the reactionmay be carried out by treating the compound of formula 9 with a compoundof formula R₂—SnA₃, where A represents a lower alkyl group such asmethyl or n-butyl, in a convenient inert solvent such as a polar aproticsolvent (e.g., N,N-dimethylformamide) or an aromatic hydrocarbon (e.g.,toluene) or acetonitrile or dimethoxyethane, in the presence of acatalytic amount of a palladium catalyst such astetrakis(triphenylphosphine)palladium(0) orbis(triphenylphosphine)palladium(II) chloride orbis(acetato)bis(triphenylphosphine)palladium(0) ortris(dibenzylideneacetone)dipalladium(0) at a temperature between about80° C. and about 180° C.

Compounds of formula 10 in which R2 represents a carboxamide or esterfunctionality with the carbonyl carbon attached to the indole ring maybe conveniently prepared using a transition metal-catalyzedcarbonylative coupling reaction. According to this process, the compoundof formula 9 is heated with an amine (to give a carboxamide product) ora lower alcohol (to give a carboxylate ester product) in the presence ofcarbon monoxide gas and a catalytic amount of a palladium catalyst suchas tetrakis(triphenylphosphine)palladium(0) or a combination of apalladium catalyst such as bis(benzonitrile)palladium(II) dichloride,with a ligand such as 1,1′-bis(diphenylphosphino)ferrocene in a solventsuch as tetrahydrofuran or toluene in a sealed tube at a temperaturebetween about 80° C. and about 100° C. Examples for specific conditionsthat may be used for such a reaction may be found in the literature, forexample in Kumar, K. et al. Org. Letters 2004, 6, 7-10.

Compounds of formula 10 in which R2 represents an aralkyl group such asa benzyl group or a substituted benzyl group may be convenientlyprepared using a transition metal-catalyzed coupling reaction such as aNegishi reaction. According to this process, the compound of formula 9is heated with an aralkyizinc reagent in the presence of a catalyticamount of a palladium catalyst such as palladium(II) acetate with aligand such as 2-dicyclohexylphosphino-2′,6′-dimethoxybiphenyl (S-Phos)or using tetrakis(triphenylphosphine)-palladium(0) in a solvent such astetrahydrofuran or toluene at a temperature between about 50° C. andabout 90° C. Examples for specific conditions that may be used for sucha reaction may be found in the literature, for example in Ellsworth, B.A. et al. US 20110082165 page 51.

Compounds of formula 10 in which R2 represents a cyanomethyl group maybe conveniently prepared using a transition metal-catalyzed couplingreaction with tributylstannanyl-acetonitrile or a lower alkylcyanoacetate. The reaction with tributylstannanyl-acetonitrile may becarried out by treating the compound of formula 9 withtributylstannanyl-acetonitrile in the presence of a palladium catalystsuch as bis(triphenylphosphino)palladium(II) dichloride orbis(tri-o-tolylphosphine)palladium(II) dichloride in toluene or xyleneat a temperature between about 110° C. and about 130° C. Conditions forsuch as reaction may be found in the literature, for example inEttaoussi, M. et al. Eur. J. Med. Chem. 2012, 49, 310-323 and in Song,D. et al. WO 2011117211 page 118. The reaction with a lower alkylcyanoacetate may be carried out by treating the compound of formula 9with the lower alkyl cyanoacetate in the presence of a palladiumcatalyst such as bis(tri-tert-butylphosphine)palladium(0), or a mixtureof a palladium catalyst such as tris-(dibenzyldeneacetone)dipalladium(0)or bis(dibenzylideneacetone)palladium(0) and a ligand such astert-butylphosphine, and a base such as trisodium phosphate in a solventsuch as toluene in a sealed tube at a temperature between about 70° C.and about 100° C. In the case where the lower alkyl cyanoacetate istest-butyl cyanoacetate, the tert-butyl group may undergo adecarboxylation reaction under the reaction conditions to give thedesired compound of formula 10 where R2 represents a cyanomethyl group.In the case where the lower alkyl cyanoacetate is methyl cyanoacetate orethyl cyanoacetate, an additional hydrolysis step is required and thisis conveniently affected by heating the product of thepalladium-catalyzed coupling with 3 M hydrochloric acid indimethylsulfoxide at about 70° C. for several hours. Examples ofconditions that may be used for such a reaction can be found in theliterature, for example in Alargova, R. G. et al. US 20120015999 page10.

The conversion of the compound of formula 10 to the compound of theinvention of formula 11 may be effected using any conventionalprocedure. For example, the reaction may be carried out by treating thecompound of formula 10 with a mixture of a base such as cesium carbonateand a lower alcohol such as methanol in a solvent such a tetrahydrofuranat a temperature between about room temperature and about the refluxtemperature of the mixture. Examples of conditions that may be used forsuch a reaction can be found in the literature, for example in Zhang, Band Wee. A. G. H. Org. Biomol. Chem. 2012, 10, 4597-4608 SupplementaryInformation; in Alain, M. et al. US 20110071150 page 54; and in Taka, N.et al. US 20120208811 Page 55.

It will be readily apparent to one of average skill in the art oforganic synthesis that, as outlined in Scheme 2, many compounds offormula 11 are also readily accessible if the protective group isremoved from the compound of formula 9 rather than from the compound offormula 10.

According to this process, the deprotection reaction may be carried outby treating the compound of formula 9 with a mixture of a base such ascesium carbonate and a lower alcohol such as methanol in a solvent sucha tetrahydrofuran at a temperature between about room temperature andabout the reflux temperature of the mixture. Examples of conditions thatmay be used for such a reaction can be found in the literature, forexample in Zhang, B and Wee. A. G. H. Org. Biomol. Chem. 2012, 10,4597-4608 Supplementary Information; in Alam, M. et al. US 20110071150page 54; and in Taka, N. et al. US 20120208811 Page 55.

The resulting compound of formula 12 may then be treated with a compoundof formula R₂—B(OH)₂, under the conditions described above for theSuzuki reaction of the compound of formula 9 to give the desiredcompound of formula 11.

Compounds of formula 11 in which R2 represents an N-linked heterocycle,such as pyrazol-1-yl may also be conveniently prepared using Scheme 2.According to this process, the compound of formula 12 is treated withthe N-linked heterocycle, such as pyrazole, in the presence of a basesuch as potassium carbonate, in the presence of a copper catalyst suchas copper(I) iodide and in the presence of L-proline, in an inertsolvent such as dimethylsulfoxide at a temperature between about 100°C., and about 130° C. Examples of conditions that may be used for such areaction can be found in the literature, for example in Sun, X. et al.Bioorg. Med. Chem. Lett. 2011, 21, 3671-3675 Supplemental Information;and in Yokotani, J. et al. US 20110275797 page 31.

Compounds of formula 21 where R3 represents a secondary amino group suchas a dialkylamino (e.g., dimethylamino diethylamino) or a cyclicsecondary amino group such as pyrrolidino, piperidino, morpholin-4-yl,1-methyl-piperazin-4-yl or the like may be prepared using the processoutlined in Scheme 3. According to this process,1-benzenesulfonyl-1H-indole-4-methanol, the compound of formula 13(which is a known compound which may be prepared according to theprocedure outlined in Castro Pineiro, J. L. et al. U.S. Pat. No.6,187,805 Column 15, or using the procedures described in the Examplesbelow) is converted to the corresponding chloromethylindole derivativeof formula 14. This undergoes a substitution reaction with an amine togive the compound of formula 15. A sequence of carboxylation,esterification, and reaction with the anion of acetonitrile, then givesthe cyanoacetyl derivative of formula 18. Reaction withdimethylformamide dimethyl acetal gives the acrylonitrile derivative offormula 19 which undergoes reaction with the arylhydrazine of formula 8to give the aminopyrazole derivative of formula 20. Removal of thephenylsulfonyl protective group then gives the compound of the inventionof formula 21.

The compound of formula 13 may be treated with methanesulfonyl chloridein the presence of a base such as triethylamine or diisopropylethylaminein a solvent such as tetrahydrofuran at a temperature about roomtemperature to give the compound of formula 14.

The compound of formula 14 may be treated with a secondary amine such asa dialkylamine (e.g., dimethylamine hydrochloride or diethylamine) or acyclic secondary amino group such as pyrrolidine, piperidine,morpholine, or 1-methyl-piperazine, in the presence of an inorganic basesuch as potassium carbonate or cesium carbonate in an inert solvent suchas acetonitrile at a temperature between about 50° C. and about 80° C.to give the amine of formula 15.

The compound of formula 15 may then be treated with a strong base suchas lithium diisopropylamide or lithium hexamethyldisilazide intetrahydrofuran at low temperature, such as at about −78° C., and thecorresponding anion treated with excess solid carbon dioxide to give thecarboxylic acid of formula 16.

The conversion of the carboxylic acid of formula 16 to the methyl esterof formula 17 may be effected using one of a variety of methods that arewell known to one of average skill in the art of organic synthesis. Manysuitable approaches are enumerated in Greene's Protective Groups inOrganic Synthesis [Wuts P. G. M and Greene, T. W., 4th Edition,Wiley-Interscience, New York, 2006, pages 553 et seq.] For example, thetransformation may be conveniently carried out by treating thecarboxylic acid of formula 16 with a chlorinating agent such as thionylchloride either neat or in an inert solvent such as benzene at atemperature between about 50° C. and about the reflux temperature. Theresulting acid chloride may then be treated with methanol in thepresence of a base such as triethylamine or diisopropylethylamine orpyridine either using methanol as solvent or in an inert solvent such astetrahydrofuran at about room temperature to give the ester of formula17.

The compound of formula 17 may be conveniently converted to thecyanoacetyl derivative of formula 18 by treating it with a mixture ofacetonitrile and a strong base such as lithium diisopropylamide orlithium hexamethyldisilazide in a solvent such as tetrahydrofuran at lowtemperature, such as at about −78° C. Conditions for such a reaction maybe found in the patent literature, for example in Taka, N. et al. US20120208811 Page 163.

The compound of formula 18 may be converted to the acrylonitrilederivative of formula 19 by treatment with N,N-dimethylformamidedimethyl acetal in an inert solvent such as an aromatic hydrocarbon(e.g., toluene) or tetrahydrofuran at about room temperature. Conditionsfor such a reaction may be found in the patent literature, for examplein Taka, N. et al. US 20120208811 page 163.

The acrylonitrile derivative of formula 19 may be converted to theaminopyrazole derivative of formula 20 by treatment with an intermediateof formula 8, where R1 is as described above in the genus of formula I,in an alcoholic solvent such as methanol or ethanol or isopropanol, atabout the reflux temperature of the solvent. Conditions for such areaction may be found in the patent literature, for example in Taka, N.et al. US 20120208811 Page 94.

The conversion of the compound of formula 20 to the compound of theinvention of formula 21 may be effected using any conventionalprocedure. For example, the reaction may be carried out by treating thecompound of formula 20 with a mixture of a base such as cesium carbonateand a lower alcohol such as methanol in a solvent such a tetrahydrofuranat a temperature between about room temperature and about the refluxtemperature of the mixture. Examples of conditions that may be used forsuch a reaction can be found in the literature, for example in Zhang, Band Wee, A. G. H. Org. Biomol. Chem. 2012, 10, 4597-4608 SupplementaryInformation; in Alam, M. et al. US 20110071150 page 54; and in Taka, N.et al. US 20120208811 Page 55.

Intermediates of formula 8 where R1 is as described above in the genusof formula I, may be prepared according to scheme 4. The compound offormula 22, 4-chloro-2-methyl-1-nitro-benzene undergoes a nucleophilicaromatic substitution reaction with a phenol derivative of formula 23 togive a compound of formula 24. Reduction of the nitro group in thecompound of formula 24, followed by diazotization and reduction givesthe aryl-hydrazine derivative of formula 8.

4-Chloro-2-methyl-1-nitro-benzene (22) may be treated with a phenol offormula 23 in the presence of a base such as potassium carbonate orcesium carbonate in an inert solvent such as dimethylformamide at atemperature between about 100° C. and about 150° C., optionally undermicrowave irradiation, to give a nitro compound of formula 24. Examplesof particular conditions that may be used for such a reaction may befound in the literature, for example in Chee, G.-L et al. US 20040266738Page 5; and in Cui, S.-L. et al. Synlett 2004, 1829-1831.

The reduction of the nitro group in the compound of formula 24 can beeffected using a variety of procedures well known to one of averageskill in the field of organic synthesis. Many of these procedures areoutlined in Larock, R. C. Comprehensive Organic Transformations JohnWiley & Sons Inc. NY 1999, pp. 823 et seq. One convenient approach is totreat the compound of formula 24 with hydrogen gas in the presence of anoble metal catalyst such as palladium-on-carbon in a solvent such analcohol (e.g., methanol or ethanol) at a pressure between about oneatmosphere of hydrogen and about three atmospheres of hydrogen at aboutroom temperature. Examples of particular conditions that may be used forsuch a reaction may be found in the literature, for example in Chee,G.-L et al. US 20040266738 Page 5; and in Schoenafinger, K. et al. US20030236288 Page 18.

The diazotization and reduction of the aniline group in the compound offormula 25 may be carried out using any conventional procedure. Forexample, the reaction is conveniently carried out by treating thecompound of formula 25 with sodium nitrite in aqueous solution in thepresence of an inorganic acid such as hydrochloric acid at a temperaturebelow about 5° C. and preferably below about 0° C., followed by theaddition of a reducing agent such as tin(II) chloride or sodiumdithionite at about the same temperature. Examples of particularconditions that may be used for such a reaction may be found in theliterature, for example in Wipf, P. and Qiming, J. WO 2012078859 page47; in Rewolinski, M. V. et al. WO 2009055721 page 82; and inSchoenafinger, K. et al. US 20030236288 page 18.

Pharmaceutical Compositions and Administration

The compounds of the present invention may be formulated in a widevariety of oral administration dosage forms and carriers. Oraladministration can be in the form of tablets, coated tablets, dragées,hard and soft gelatin capsules, solutions, emulsions, syrups, orsuspensions. Compounds of the present invention are efficacious whenadministered by other routes of administration including continuous(intravenous drip) topical parenteral, intramuscular, intravenous,subcutaneous, transdermal (which may include a penetration enhancementagent), buccal, nasal, inhalation and suppository administration, amongother routes of administration. The preferred manner of administrationis generally oral using a convenient daily dosing regimen which can beadjusted according to the degree of affliction and the patient'sresponse to the active ingredient.

A compound or compounds of the present invention, as well as theirpharmaceutically useable salts, together with one or more conventionalexcipients, carriers, or diluents, may be placed into the form ofpharmaceutical compositions and unit dosages. The pharmaceuticalcompositions and unit dosage forms may be comprised of conventionalingredients in conventional proportions, with or without additionalactive compounds or principles, and the unit dosage forms may containany suitable effective amount of the active ingredient commensurate withthe intended daily dosage range to be employed. The pharmaceuticalcompositions may be employed as solids, such as tablets or filledcapsules, semisolids, powders, sustained release formulations, orliquids such as solutions, suspensions, emulsions, elixirs, or filledcapsules for oral use; or in the form of suppositories for rectal orvaginal administration; or in the form of sterile injectable solutionsfor parenteral use. A typical preparation will contain from about 5% toabout 95% active compound or compounds (w/w). The term “preparation” or“dosage form” is intended to include both solid and liquid formulationsof the active compound and one skilled in the art will appreciate thatan active ingredient can exist in different preparations depending onthe target organ or tissue and on the desired dose and pharmacokineticparameters.

The term “excipient” as used herein refers to a compound that is usefulin preparing a pharmaceutical composition, generally safe, non-toxic andneither biologically nor otherwise undesirable, and includes excipientsthat are acceptable for veterinary use as well as human pharmaceuticaluse. The compounds of this invention can be administered alone but willgenerally be administered in admixture with one or more suitablepharmaceutical excipients, diluents or carriers selected with regard tothe intended route of administration and standard pharmaceuticalpractice.

“Pharmaceutically acceptable” means that which is useful in preparing apharmaceutical composition that is generally safe, non-toxic, andneither biologically nor otherwise undesirable and includes that whichis acceptable for veterinary as well as human pharmaceutical use.

A “pharmaceutically acceptable salt” form of an active ingredient mayalso initially confer a desirable pharmacokinetic property on the activeingredient which were absent in the non-salt form, and may evenpositively affect the pharmacodynamics of the active ingredient withrespect to its therapeutic activity in the body. The phrase“pharmaceutically acceptable salt” of a compound means a salt that ispharmaceutically acceptable and that possesses the desiredpharmacological activity of the parent compound. Such salts include: (1)acid addition salts, formed with inorganic acids such as hydrochloricacid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, andthe like; or formed with organic acids such as acetic acid, propionicacid, hexanoic acid, cyclopentanepropionic acid, glycolic acid, pyruvicacid, lactic acid, malonic acid, succinic acid, malic acid, maleic acid,fumaric acid, tartaric acid, citric acid, benzoic acid,3-(4-hydroxybenzoyl)benzoic acid, cinnamic acid, mandelic acid,methanesulfonic acid, ethanesulfonic acid, 1,2-ethane-disulfonic acid,2-hydroxyethanesulfonic acid, benzenesulfonic acid,4-chlorobenzenesulfonic acid, 2-naphthalenesulfonic acid,4-toluenesulfonic acid, camphorsulfonic acid,4-methylbicyclo[2.2.2]-oct-2-ene-1-carboxylic acid, glucoheptonic acid,3-phenylpropionic acid, trimethylacetic acid, tertiary butylacetic acid,lauryl sulfuric acid, gluconic acid, glutamic acid, hydroxynaphthoicacid, salicylic acid, stearic acid, muconic acid, and the like; or (2)salts formed when an acidic proton present in the parent compound eitheris replaced by a metal ion, e.g., an alkali metal ion, an alkaline earthion, or an aluminum ion; or coordinates with an organic base such asethanolamine, diethanolamine, triethanolamine, tromethamine,N-methylglucamine and the like.

Solid form preparations include powders, tablets, pills, capsules,cachets, suppositories, and dispersible granules. A solid carrier may beone or more substances which may also act as diluents, flavoring agents,solubilizers, lubricants, suspending agents, hinders, preservatives,tablet disintegrating agents, or an encapsulating material. In powders,the carrier generally is a finely divided solid which is a mixture withthe finely divided active component. In tablets, the active componentgenerally is mixed with the carrier having the necessary bindingcapacity in suitable proportions and compacted in the shape and sizedesired. Suitable carriers include but are not limited to magnesiumcarbonate, magnesium stearate, talc, sugar, lactose, pectin, dextrin,starch, gelatin, tragacanth, methylcellulose, sodiumcarboxymethylcellulose, a low melting wax, cocoa butter, and the like.Solid form preparations may contain, in addition to the activecomponent, colorants, flavors, stabilizers, buffers, artificial andnatural sweeteners, dispersants, thickeners, solubilizing agents, andthe like.

Liquid formulations also are suitable for oral administration includeliquid formulation including emulsions, syrups, elixirs, aqueoussolutions, aqueous suspensions. These include solid form preparationswhich are intended to be converted to liquid form preparations shortlybefore use. Emulsions may be prepared in solutions, for example, inaqueous propylene glycol solutions or may contain emulsifying agentssuch as lecithin, sorbitan monooleate, or acacia. Aqueous solutions canbe prepared by dissolving the active component in water and addingsuitable colorants, flavors, stabilizing, and thickening agents. Aqueoussuspensions can be prepared by dispersing the finely divided activecomponent in water with viscous material, such as natural or syntheticgums, resins, methylcellulose, sodium carboxymethylcellulose, and otherwell known suspending agents.

The compounds of the present invention may be formulated for parenteraladministration (e.g., by injection, for example bolus injection orcontinuous infusion) and may be presented in unit dose form in ampoules,pre-filled syringes, small volume infusion or in multi-dose containerswith an added preservative. The compositions may take such forms assuspensions, solutions, or emulsions in oily or aqueous vehicles, forexample solutions in aqueous polyethylene glycol. Examples of oily ornonaqueous carriers, diluents, solvents or vehicles include propyleneglycol, polyethylene glycol, vegetable oils (e.g., olive oil), andinjectable organic esters (e.g., ethyl oleate), and may containformulatory agents such as preserving, wetting, emulsifying orsuspending, stabilizing and/or dispersing agents. Alternatively, theactive ingredient may be in powder form, obtained by aseptic isolationof sterile solid or by lyophilization from solution for constitutionbefore use with a suitable vehicle, e.g., sterile, pyrogen-free water.

The compounds of the present invention may be formulated for topicaladministration to the epidermis as ointments, creams or lotions, or as atransdermal patch. Ointments and creams may, for example, be formulatedwith an aqueous or oily base with the addition of suitable thickeningand/or gelling agents. Lotions may be formulated with an aqueous or oilybase and will in general also containing one or more emulsifying agents,stabilizing agents, dispersing agents, suspending agents, thickeningagents, or coloring agents. Formulations suitable for topicaladministration in the mouth include lozenges comprising active agents ina flavored base, usually sucrose and acacia or tragacanth; pastillescomprising the active ingredient in an inert base such as gelatin andglycerin or sucrose and acacia; and mouthwashes comprising the activeingredient in a suitable liquid carrier.

The compounds of the present invention may be formulated foradministration as suppositories. A low melting wax, such as a mixture offatty acid glycerides or cocoa butter is first melted and the activecomponent is dispersed homogeneously, for example, by stirring. Themolten homogeneous mixture is then poured into convenient sized molds,allowed to cool, and to solidify.

The compounds of the present invention may be formulated far vaginaladministration. Pessaries, tampons, creams, gels, pastes, foams orsprays containing in addition to the active ingredient such carriers asare known in the art to be appropriate.

The compounds of the present invention may be formulated for nasaladministration. The solutions or suspensions are applied directly to thenasal cavity by conventional means, for example, with a dropper, pipetteor spray. The formulations may be provided in a single or multidoseform. In the latter case of a dropper or pipette, this may be achievedby the patient administering an appropriate, predetermined volume of thesolution or suspension. In the case of a spray, this may be achieved forexample by means of a metering atomizing spray pump.

The compounds of the present invention may be formulated for aerosoladministration, particularly to the respiratory tract and includingintranasal administration. The compound will generally have a smallparticle size for example of the order of five (5) microns or less. Sucha particle size may be obtained by means known in the art, for exampleby micronization. The active ingredient is provided in a pressurizedpack with a suitable propellant such as a chlorofluorocarbon (CFC), forexample, dichlorodifluoromethane, trichlorofluoromethane, ordichlorotetrafluoroethane, or carbon dioxide or other suitable gas. Theaerosol may conveniently also contain a surfactant such as lecithin. Thedose of drug may be controlled by a metered valve. Alternatively theactive ingredients may be provided in a form of a dry powder, forexample a powder mix of the compound in a suitable powder base such aslactose, starch, starch derivatives such as hydroxypropylmethylcellulose and polyvinylpyrrolidine (PVP). The powder carrier will form agel in the nasal cavity. The powder composition may be presented in unitdose form for example in capsules or cartridges of e.g., gelatin orblister packs from which the powder may be administered by means of aninhaler.

When desired, formulations can be prepared with enteric coatings adaptedfor sustained or controlled release administration of the activeingredient. For example, the compounds of the present invention can beformulated in transdermal or subcutaneous drug delivery devices. Thesedelivery systems are advantageous when sustained release of the compoundis necessary and when patient compliance with a treatment regimen iscrucial. Compounds in transdermal delivery systems are frequentlyattached to an skin-adhesive solid support. The compound of interest canalso be combined with a penetration enhancer, e.g., Azone(1-dodecylaza-cycloheptan-2-one). Sustained release delivery systems areinserted subcutaneously into to the subdermal layer by surgery orinjection. The subdermal implants encapsulate the compound in a lipidsoluble membrane, e.g., silicone rubber, or a biodegradable polymer,e.g., polylactic acid.

Suitable formulations along with pharmaceutical carriers, diluents andexcipients are described in Remington: The Science and Practice ofPharmacy 1995, edited by E. W. Martin, Mack Publishing Company, 19thedition, Easton, Pa. A skilled formulation scientist may modify theformulations within the teachings of the specification to providenumerous formulations for a particular route of administration withoutrendering the compositions of the present invention unstable orcompromising their therapeutic activity.

The modification of the present compounds to render them more soluble inwater or other vehicle, for example, may be easily accomplished by minormodifications (salt formulation, esterification, etc.), which are wellwithin the ordinary skill in the art. It is also well within theordinary skill of the art to modify the route of administration anddosage regimen of a particular compound in order to manage thepharmacokinetics of the present compounds for maximum beneficial effectin patients.

The term “therapeutically effective amount” as used herein means anamount required to reduce symptoms of the disease in an individual. Thedose will be adjusted to the individual requirements in each particularcase. That dosage can vary within wide limits depending upon numerousfactors such as the severity of the disease to be treated, the age andgeneral health condition of the patient, other medicaments with whichthe patient is being treated, the route and form of administration andthe preferences and experience of the medical practitioner involved. Fororal administration, a daily dosage of between about 0.01 and about 1000mg/kg body weight per day should be appropriate in monotherapy and/or incombination therapy. A preferred daily dosage is between about 0.1 andabout 500 mg/kg body weight, more preferred 0.1 and about 100 mg/kg bodyweight and most preferred 1.0 and about 10 mg/kg body weight per day.Thus, for administration to a 70 kg person, the dosage range would beabout 7 mg to 0.7 g per day. The daily dosage can be administered as asingle dosage or in divided dosages, typically between 1 and 5 dosagesper day. Generally, treatment is initiated with smaller dosages whichare less than the optimum dose of the compound. Thereafter, the dosageis increased by small increments until the optimum effect for theindividual patient is reached. One of ordinary skill in treatingdiseases described herein will be able, without undue experimentationand in reliance on personal knowledge, experience and the disclosures ofthis application, to ascertain a therapeutically effective amount of thecompounds of the present invention for a given disease and patient.

The pharmaceutical preparations are preferably in unit dosage forms. Insuch form, the preparation is subdivided into unit doses containingappropriate quantities of the active component. The unit dosage form canbe a packaged preparation, the package containing discrete quantities ofpreparation, such as packeted tablets, capsules, and powders in vials orampoules. Also, the unit dosage form can be a capsule, tablet, cachet,or lozenge itself, or it can be the appropriate number of any of thesein packaged form.

Indications and Methods of Treatment

The compounds of generic Formula I inhibit Bruton's tyrosine kinase(Btk). Activation of Btk by upstream kinases results in activation ofphospholipase-Cγ which, in turn, stimulates release of pro-inflammatorymediators. Compounds of Formula I are useful in the treatment ofarthritis and other anti-inflammatory and auto-immune diseases.Compounds according to Formula I are, accordingly, useful for thetreatment of arthritis. Compounds of Formula I are useful for inhibitingBtk in cells and for modulating B-cell development. The presentinvention further comprises pharmaceutical compositions containingcompounds of Formula I admixed with pharmaceutically acceptable carrier,excipients or diluents.

The compounds described herein are kinase inhibitors, in particular Btkinhibitors. These inhibitors can be useful for treating one or morediseases responsive to kinase including diseases responsive to Btkinhibition and/or inhibition of B-cell proliferation, in mammals.Without wishing to be bound to any particular theory, it is believedthat the interaction of the compounds of the invention with Btk resultsin the inhibition of Btk activity and thus in the pharmaceutical utilityof these compounds. Accordingly, the invention includes a method oftreating a mammal, for instance a human, having a disease responsive toinhibition of Btk activity, and/or inhibiting B-cell proliferation,comprising administrating to the mammal having such a disease, aneffective amount of at least one chemical entity provided herein. Aneffective concentration may be ascertained experimentally, for exampleby assaying blood concentration of the compound, or theoretically, bycalculating bioavailability. Other kinases that may be affected inaddition to Btk include, but are not limited to, other tyrosine kinasesand serine/threonine kinases.

Kinases play notable roles in signaling pathways controlling fundamentalcellular processes such as proliferation, differentiation, and death(apoptosis). Abnormal kinase activity has been implicated in a widerange of diseases, including multiple cancers, autoimmune and/orinflammatory diseases, and acute inflammatory reactions. Themultifaceted role of kinases in key cell signaling pathways provides asignificant opportunity to identify novel drugs targeting kinases andsignaling pathways.

An embodiment includes a method of treating a patient having anautoimmune and/or inflammatory disease, or an acute inflammatoryreaction responsive to inhibition of Btk activity and/or B-cellproliferation.

Autoimmune and/or inflammatory diseases that can be affected usingcompounds and compositions according to the invention include, but arenot limited to: psoriasis, allergy, Crohn's disease, irritable bowelsyndrome, Sjogren's disease, tissue graft rejection, and hyperacuterejection of transplanted organs, asthma, systemic lupus erythematosus(and associated glomerulonephritis), dermatomyositis, multiplesclerosis, scleroderma, vasculitis (ANCA-associated and othervasculitides), autoimmune hemolytic and thrombocytopenic states,Goodpasture's syndrome (and associated glomerulonephritis and pulmonaryhemorrhage), atherosclerosis, rheumatoid arthritis, chronic Idiopathicthrombocytopenic purpura (ITP), Addison's disease, Parkinson's disease,Alzheimer's disease, diabetes, septic shock, and myasthenia gravis.

Included herein are methods of treatment in which at least one chemicalentity provided herein is administered in combination with ananti-inflammatory agent. Anti-inflammatory agents include but are notlimited to NSAIDs, non-specific and COX-2 specific cyclooxgenase enzymeinhibitors, gold compounds, corticosteroids, methotrexate, tumornecrosis factor receptor (TNF) receptors antagonists, immunosuppressantsand methotrexate.

Examples of NSAIDs include, but are not limited to, ibuprofen,flurbiprofen, naproxen and naproxen sodium, diclofenac, combinations ofdiclofenac sodium and misoprostol, sulindac, oxaprozin, diflunisal,piroxicam, indomethacin, etodolac, fenoprofen calcium, ketoprofen,sodium nabumetone, sulfasalazine, tolmetin sodium, andhydroxychloroquine. Examples of NSAIDs also include COX-2 specificinhibitors such as celecoxib, valdecoxib, lumiracoxib and/or etoricoxib.

In some embodiments, the anti-inflammatory agent is a salicylate.Salicylates include by are not limited to acetylsalicylic acid oraspirin, sodium salicylate, and choline and magnesium salicylates.

The anti-inflammatory agent may also be a corticosteroid. For example,the corticosteroid may be cortisone, dexamethasone, methylprednisolone,prednisolone, prednisolone sodium phosphate, or prednisone.

In additional embodiments the anti-inflammatory agent is a gold compoundsuch as gold sodium thiomalate or auranofin.

The invention also includes embodiments in which the anti-inflammatoryagent is a metabolic inhibitor such as a dihydrofolate reductaseinhibitor, such as methotrexate or a dihydroorotate dehydrogenaseinhibitor, such as leflunomide.

Other embodiments of the invention pertain to combinations in which atleast one anti-inflammatory compound is an anti-C5 monoclonal antibody(such as eculizumab or pexelizumab), a TNF antagonist, such asentanercept, or infliximab, which is an anti-TNF alpha monoclonalantibody.

Still other embodiments of the invention pertain to combinations inwhich at least one active agent is an iminunosuppressant compound suchas an immunosuppressant compound chosen from methotrexate, leflunomide,cyclosporine, tacrolimus, azathioprine, and mycophenolate mofetil.

B-cells and B-cell precursors expressing BTK have been implicated in thepathology of B-cell malignancies, including, but not limited to, B-celllymphoma, lymphoma (including Hodgkin's and non-Hodgkin's lymphoma),hairy cell lymphoma, multiple myeloma, chronic and acute myelogenousleukemia and chronic and acute lymphocytic leukemia.

BTK has been shown to be an inhibitor of the Fas/APO-1 (CD-95) deathinducing signaling complex (DISC) in B-lineage lymphoid cells. The fateof leukemia/lymphoma cells may reside in the balance between theopposing proapoptotic effects of caspases activated by DISC and anupstream anti-apoptotic regulatory mechanism involving BTK and/or itssubstrates (Vassilev et al., J. Biol. Chem. 1998, 274, 1646-4656).

It has also been discovered that BTK inhibitors are useful aschemosensitizing agents, and, thus, are useful in combination with otherchemotherapeutic drugs, in particular, drugs that induce apoptosis.Examples of other chemotherapeutic drugs that can be used in combinationwith chemosensitizing BTK inhibitors include topoisomerase I inhibitors(camptothecin or topotecan), topoisomerase II inhibitors (e.g.daunomycin and etoposide), alkylating agents (e.g. cyclophosphamide,melphalan and BCNU), tubulin directed agents (e.g. taxol andvinblastine), and biological agents (e.g. antibodies such as anti CD20antibody, IDEC 8, immunotoxins, and cytokines).

Btk activity has also been associated with some leukemias expressing thebcr-abl fusion gene resulting from translocation of parts of chromosome9 and 22. This abnormality is commonly observed in chronic myelogenousleukemia. Btk is constitutively phosphorylated by the bcr-abl kinasewhich initiates downstream survival signals which circumvents apoptosisin bcr-abl cells. (N. Feldhahn et al. J. Exp. Med. 2005201(10:1837-1852).

Methods of Treatment

The application provides a method for treating an inflammatory and/orautoimmune condition comprising administering to a patient in needthereof a therapeutically effective amount of the compound of Formula I.

The application provides a method for treating an inflammatory conditioncomprising administering to a patient in need thereof a therapeuticallyeffective amount of the compound of Formula I.

The application provides a method for treating rheumatoid arthritiscomprising administering to a patient in need thereof a therapeuticallyeffective amount of the compound of Formula 1.

The application provides a method for treating asthma comprisingadministering to a patient in need thereof a therapeutically effectiveamount of Formula I.

The application provides a method for treating an inflammatory and/orautoimmune condition comprising administering to a patient in needthereof a therapeutically effective amount of the Btk inhibitor compoundof Formulae I.

The application provides a method for treating arthritis comprisingadministering to a patient need thereof a therapeutically effectiveamount of the Btk inhibitor compound of Formula I.

The application provides a method for treating asthma comprisingadministering to a patient in need thereof a therapeutically effectiveamount of the Btk inhibitor compound of Formula I.

The application provides a method for treating cancer comprisingadministering to a patient in need thereof a therapeutically effectiveamount of the Btk inhibitor compound of Formula I.

The application provides a method of inhibiting B-cell proliferationcomprising administering to a patient in need thereof a therapeuticallyeffective amount of the Btk inhibitor compound of Formula I.

The application provides a method for inhibiting Btk activity comprisingadministering the Btk inhibitor compound of any one of Formula I,wherein the Btk inhibitor compound exhibits an IC₅₀ of 50 micromolar orless in an in vitro biochemical assay of Btk activity.

In one variation of the above method, the Btk inhibitor compoundexhibits an IC₅₀ of 100 nanomolar or less in an in vitro biochemicalassay of Btk activity.

In another variation of the above method, the compound exhibits an IC₅₀of 10 nanomolar or less in an in vitro biochemical assay of Btkactivity.

The application provides a method for treating an inflammatory conditioncomprising co-administering to a patient in need thereof atherapeutically effective amount of an anti-inflammatory compound incombination with the Btk inhibitor compound of Formula I.

The application provides a method for treating arthritis comprisingco-administering to a patient in need thereof a therapeuticallyeffective amount of an anti-inflammatory compound in combination withthe Btk inhibitor compound of Formula 1.

The application provides a method for treating a lymphoma or a BCR-ABL1⁺leukemia cells by administering to a patient in need thereof atherapeutically effective amount of the Btk inhibitor compound ofFormula I.

EXAMPLES

General Abbreviations

Commonly used abbreviations include: acetyl (Ac),azo-bis-isobutyrylnitrile (AIBN), atmospheres (Atm),9-borabicyclo[3.3.1]nonane (9-BBN or BBN),2,2′-bis(diphenylphosphino)-1,1′-binaphthyl (BINAP), tert-butoxycarbonyl(Boc), di-tert-butyl pyrocarbonate or boc anhydride (BOC₂O), benzyl(Bn), butyl (Bu), Chemical Abstracts Registration Number (CASRN),benzyloxycarbonyl (CBZ or Z), carbonyl diimidazole (CDI),1,4-diazabicyclo[2.2.2]octane (DABCO), diethylaminosulfur trifluoride(DAST), dibenzylideneacetone (dba), 1,5-diazabicyclo[4.3.0]non-5-enc(DBN), 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU),N,N′-dicyclohexylcarbodiimide (DCC), 1,2-dichloroethane (DCE),dichloromethane (DCM), 2,3-Dichloro-5,6-dicyano-1,4-benzoquinone (DDQ),diethyl azodicarboxylate (DEAD), di-iso-propylazodicarboxylate (DIAD),di-iso-butylaluminumhydride (DIBAL or DIBAL-H), di-iso-propylethylamine(DIPEA), N,N-dimethyl acetamide (DMA), 4-N,N-dimethylaminopyridine(DMAP), N,N-dimethylformamide (DMF), dimethyl sulfoxide (DMSO),1,1′-bis-(diphenylphosphino)ethane (dppe),1,1′-bis-(diphenylphosphino)ferrocene (dppf),1-(3-dimethylaininopropyl)-3-ethylcarbodiimide hydrochloride (EDCI),2-ethoxy-1-ethoxycarbonyl-1,2-dihydroquinoline (EEDQ), ethyl (Et), ethylacetate (EtOAc), ethanol (EtOH), 2-ethoxy-2H-quinoline-1-carboxylic acidethyl ester (EEDQ), diethyl ether (Et₂O), ethyl isopropyl ether(EtOiPr), O-(7-azabenzotriazole-1-yl)-N, N,N′,N′-tetramethyluroniumhexafluorophosphate acetic acid (HATU), acetic acid (HOAc),1-N-hydroxybenzotriazole (HOBt), high pressure liquid chromatography(HPLC), iso-propanol (IPA), isopropylmagnesium chloride (iPrMgCl),hexamethyl disilazane (HMDS), liquid chromatography mass spectrometry(LCMS), lithium hexamethyl disilazane (LiHMDS), meta-chloroperoxybenzoicacid (m-CPBA), methanol (MeOH), melting point (mp), MeSO₂— (mesyl orMs), methyl (Me), acetonitrile (MeCN), m-chloroperbenzoic acid (MCPBA),mass spectrum (ms), methyl t-butyl ether (MTBE), methyl tetrahydrofuran(MeTHF), N-bromosuccinimide (NBS), n-Butyllithium (nBuLi),N-carboxyanhydride (NCA), N-chlorosuccinimide (NCS), N-methylmorpholine(NMM), N-methylpyrrolidone (NMP), pyridinium chlorochromate (PCC),Dichloro-((bis-diphenylphosphino)ferrocenyl) palladium(H) (Pd(dppf)Cl₂),palladium(II) acetate (Pd(OAc)₂),tris(dibenzylideneacetone)dipalladium(0) (Pd₂(dba)₃), pyridiniumdichromate (PDC), phenyl (Ph), propyl (Pr), iso-propyl (i-Pr), poundsper square inch (psi), pyridine (pyr),1,2,3,4,5-Pentaphenyl-1′-(di-tert-butylphosphino)ferrocene (Q-Phos),room temperature (ambient temperature, rt or RT), sec-Butyllithium(sBuLi), tert-butyldimethylsilyl or t-BuMe₂Si (TBDMS),tetra-n-butylammonium fluoride (TBAF), triethylamine (TEA or Et₃N),2,2,6,6-tetramethylpiperidine 1-oxyl (TEMPO), trimethylsilylethoxymethyl(SEM), triflate or CP₃SO₂-(Tf), trifluoroacetic acid (TFA),1,1′-bis-2,2,6,6-tetramethylheptane-2,6-dione (TMHD),O-benzotriazol-1-yl-N,N,N′,N′-tetramethyluronium tetrafluoroborate(TBTU), thin layer chromatography (TLC), tetrahydrofuran (THF),trimethylsilyl or Me₃Si (TMS), p-toluenesulfonic acid monohydrate (TsOHor pTsOH), 4-Me-C₆H₄SO₂— or tosyl (Ts), andN-urethane-N-carboxyanhydride (UNCA). Conventional nomenclatureincluding the prefixes normal (n), iso (i-), secondary (sec-), tertiary(tert-) and neo have their customary meaning when used with an alkylmoiety. (J. Rigaudy and D. P. Klesney, Nomenclature in OrganicChemistry, IUPAC 1979 Pergamon Press, Oxford.).

General Conditions

Compounds of the present invention cart be prepared beginning with thecommercially available starting materials by utilizing general synthetictechniques and procedures known to those skilled in the art. Outlinesbelow are reaction schemes suitable for preparing such compounds.Further exemplification can be found in the specific examples.

PREPARATIVE EXAMPLES

Specific Abbreviations

CDCl₃ Deuterated chloroform

CH₂Cl₂ Dichloromethane

CH₃CN Acetonitrile

CO₂ Carbon dioxide

Cone Concentrated

Cs₂CO₃ Cesium carbonate

DIPEA Diisopropylethylamine

DMF N,N-Dimethylformamide

DMSO Dimethylsulfoxide

EtOAc Ethyl acetate

EtOH Ethanol

HCl Hydrochloric acid

K₂CO₃ Potassium carbonate

LDA Lithium diisopropylamide

LiAlH₄ Lithium aluminum hydride

MeOH Methanol

NaBH₄ Sodium borohydride

NaOH Sodium hydroxide

Na₂SO₄ Sodium sulfate

NaH Sodium hydride

NaNO₂ Sodium nitrite

Pd(OAc)₂ Palladium(II) acetate

Pd(PPh₃)₄ Tetrakis(triphenylphosphine)palladium(0)

SOCl₂ Thionyl chloride

THE Tetrahydrofuran

General Experimental Details

Reagents were purchased from Aldrich, Oakwood, Matrix or other suppliersand used without further purification. Reactions using microwaveirradiation for heating were conducted using either a Personal ChemistryEmrys Optimizer System or a CEM Discovery System. The purification ofmulti-milligram to multi-gram scale was conducted by methods known knowto those skilled in the art such as elution of silica gel flash column;preparative flash column purifications were also effected in some casesby use of disposal pre-packed multigram silica gel columns (RediSep)eluted with a CombiFlash system. Biotage™ and ISCO™ are also flashcolumn instruments that may have been used in this invention forpurification of intermediates.

For the purpose of judging compound identity and purity, LC/MS (liquidchromatography/mass spectroscopy) spectra were recorded using thefollowing system. For measurement of mass spectra, the system consistsof a Micromass Platform II spectrometer: ES Ionization in positive mode(mass range: 150-1200). The simultaneous chromatographic separation wasachieved with the following HPLC system: ES Industries Chromegabond WRC-18 3u 120 Å (3.2×30 mm) column cartridge; Mobile Phase A: Water (0.02%TEA) and Phase B: Acetonitrile (0.02% TFA); gradient 10% B to 90% B in 3minutes; equilibration time of 1 minute; flow rate of 2 mL/minute.

Many compounds of Formula 1 were also purified by reversed phased HPLC,using methods well known to those skilled in the art. In some cases,preparative HPLC purification was conducted using PE Sciex 150 EX MassSpec controlling a Gilson 215 collector attached to a Shimadzupreparative HPLC system and a Leap autoinjector. Compounds werecollected from the elution stream using LC/MS detection in the positiveion detection: The elution of compounds from C-18 columns (2.0×10 cmeluting at 20 mL/min) was effected using appropriate linear gradationmode over 10 minutes of Solvent (A) 0.05% TFA/H₂O and Solvent (B) 0.035%TFA/acetonitrile. For injection on to HPLC systems, the crude sampleswere dissolved in mixtures of methanol, acetonitrile and DMSO.

¹H-NMR characterization was performed using Bruker or Varian 300 or 400MHz NMR Spectrometers.

The compounds of the present invention may be synthesized according toknown techniques. The following examples and references are provided toaid the understanding of the present invention. The examples are notintended, however, to limit the invention, the true scope of which isset forth in the appended claims. The names of the final products in theexamples were generated using Isis AutoNom 2000.

Preparation of Intermediates Intermediate 1[4-(2,3-Difluoro-phenoxy)-2-methyl-phenyl]-hydrazine

Step 1: 4-(2,3-Difluoro-phenoxy)-2-methyl-1-nitro-benzene

A mixture of 4-chloro-2-methyl-1-nitro-benzene (5 g, 29.1 mmol),2,3-difluorophenol (4.55 g, 35.0 mmol) and Cs₂CO₃ (14.2 g, 43.7 mmol) inDMF (10 mL) was heated in a sealed tube in a microwave oven at 150° C.for 30 min. EtOAc (300 mL) was added and the mixture was washed withwater (150 mL) and brine. The organic layer was dried (Na₂SO₄),filtered, and evaporated. The crude material was purified by flashchromatography (silica gel, 10% ethyl acetate in hexanes) to give4-(2,3-difluoro-phenoxy)-2-methyl-1-nitro-benzene (5.6 g, 72%) as alight yellow oil.

Step 2: 4-(2,3-Difluoro-phenoxy)-2-methyl-phenylamine

A mixture of 4-(2,3-difluoro-phenoxy)-2-methyl-1-nitro-benzene (5.28 g,19.9 mmol) and palladium-on-carbon (587 mg) in MeOH (55 mL) was shakenunder 30 psi of hydrogen in a Parr shaker for 4 h. The mixture wasfiltered through Celite and the Celite was washed with MeOH. Thefiltrate was evaporated to give4-(2,3-difluoro-phenoxy)-2-methyl-phenylamine (4.56 g, 97%) which wasused in the next step without further purification.

Step 3: [4-(2,3-Difluoro-phenoxy)-2-methyl-phenyl]-hydrazine

A mixture of 4-(2,3-difluoro-phenoxy)-2-methyl-phenylamine (546 mg, 2.32mmol) and conc. HCl (0.6 mL) in MeOH (1 mL) and water (2 mL) was cooledin an ice bath. A solution of NaNO₂ (168 mg, 2.44 mmol) in water (0.4mL) was added slowly and the mixture was stirred for 30 min. Thereaction mixture was transferred by pipette to a stirred solution oftin(II) chloride dihydrate (2.25 g, 10 mmol) in conc. HCl (5 mL) and themixture was stirred for 4 h. MeOH (4 mL) was added, followed by 10 MNaOH until the pH reached 7-8. The solvent was evaporated under reducedpressure and the residue was dried under vacuum for 1 h. The residue wastriturated with 10% MeOH/CH₂Cl₂, and the solid was filtered off andwashed with 10% MeOH/CH₂Cl₂. The filtrate was evaporated under reducedpressure to give [4-(2,3-difluoro-phenoxy)-2-methyl-phenyl]-hydrazine(495 mg, 68%) as a yellow oil. This material was used in the next stepwithout further purification.

Intermediate 2 3-(4-Hydrazino-3-methyl-phenoxy)-benzonitrile

A mixture of 3-(4-amino-3-methyl-phenoxy)-benzonitrile (which may beprepared as described in Akama, T. et al. Bioorg. Med. Chem. Lett. 2009,19, 2129-2132; 2.2 g, 9.8 mmol) and conc. HCl (3.5 mL) in MeOH (7 mL)and water (5 mL) was cooled in an ice bath. A solution of NaNO₂ (1.35 g,19.6 mmol) in water (6 mL) was added slowly and the mixture was stirredfor 30 min. The reaction mixture was transferred by pipette to a stirredsolution of tin(II) chloride dihydrate (8.85 g, 39.2 mmol) in conc. HCl(7 mL) and the mixture was stirred for 30 min. MeOH (10 mL) was added,followed by 10 M NaOH until the pH reached 7-8. Water (100 mL) was addedand the mixture was extracted with EtOAc (500 mL). The organic layer waswashed with brine (Na₂SO₄), filtered, and evaporated to give3-(4-hydrazino-3-methyl-phenoxy)-benzonitrile (2.2 g, 94%) as an oil.This material was used in the next step without further purification.

Example I-1{5-Amino-1-[4-(2,3-difluoro-phenoxy)-2-methyl-phenyl]-1H-pyrazol-4-yl}-(4-bromo-1H-indol-2-yl)-methanone

Step 1: 1-Benzenesulfonyl-4-bromo-1H-indole-2-carboxylic acid methylester

DIPEA (2.6 mL, 14.8 mmol) and MeOH (3 mL, 74.2 mmol) were added to asolution of 1-benzenesulfonyl-4-bromo-1H-indole-2-carbonyl chloride(which may be prepared as described in Mahboobi, S. et al, J. Med. Chem.2006, 49, 3101-3115; 1.97 g, 4.94 mmol). The mixture was stirred at roomtemperature overnight, and the solvent was evaporated under reducedpressure. The residue was triturated with 15% EtOAc/hexanes, filtered,washed with 15% EtOAc/hexanes, and dried under vacuum overnight to give1-benzenesulfonyl-4-promo-1H-indole-2-carboxylic acid methyl ester (1.78g, 91%).

Step 2: 3-(1-Benzenesulfonyl-4-bromo-1H-indol-2-yl)-3-oxo-propionitrile

A solution of methyl 4-bromo-1-(phenylsulfonyl)-1H-indole-2-carboxylate(970 mg, 2.46 mmol) and acetonitrile (770 μL, 14.8 mmol) in THF (25 mL)was cooled to −78° C. LDA (2M/THF) (2.5 ml, 5 mmol) was added slowlyover 5 min. The reaction mixture was stirred at −78° C. for 30 min andthen saturated NH₄Cl solution (40 mL) was added. Water (150 mL) wasadded and the mixture was extracted with EtOAc (500 mL). The organiclayer was washed with brine, dried (Na₂SO₄), filtered, and evaporated.The residue was purified by chromatography (silica gel, 30%EtOAc/hexanes) to give3-(4-bromo-1-(phenylsulfonyl)-1H-indol-2-yl)-3-oxopropanenitrile (650mg, 66%) as a foam.

Step 3:(E)-2-(1-Benzenesulfonyl-4-bromo-1H-indole-2-carbonyl)-3-dimethylamino-acrylonitrile

N,N-dimethylformamide dimethyl acetal (465 mg, 3.9 mmol) was added to asolution of3-(4-bromo-1-(phenylsulfonyl)-1H-indol-2-yl)-3-oxopropanenitrile (1.21g, 3.00 mmol) in toluene (20 mL), and the mixture was stirred at roomtemperature overnight. The solvent was removed under reduced pressure.The residue was purified by chromatography (silica gel, 70%EtOAc/hexanes) to give(E)-2-(1-benzenesulfonyl-4-bromo-1H-indole-2-carbonyl)-3-dimethylamino-acrylonitrile(1.06 g, 77%) as a yellow foam.

Step 4:{5-Amino-1-[4-(2,3-difluoro-phenoxy)-2-methyl-phenyl]-1H-pyrazol-4-yl}-(1-benzenesulfonyl-4-bromo-1H-indol-2-yl)-methanone

A mixture of [4-(2,3-difluoro-phenoxy)-2-methyl-phenyl]-hydrazine (whichmay be prepared as described for Intermediate 1; 816 mg, 3.26 mmol),(E)-2-(1-benzenesulfonyl-4-bromo-1H-indole-2-carbonyl)-3-dimethylamino-acrylonitrile(650 mg, 1.42 mmol) and EtOH (25 mL) was heated at reflux overnight. Thesolvent was removed under reduced pressure. The residue was purified bychromatography (silica gel, 30% EtOAc/hexanes) to give{5-amino-1-[4-(2,3-difluoro-phenoxy)-2-methyl-phenyl]-1H-pyrazol-4-yl}-(1-benzenesulfonyl-4-bromo-1H-indol-2-yl)-methanone(820 mg, 87%) as a foam.

Step 5:{5-Amino-1-[4-(1,3-difluoro-phenoxy)-2-methyl-phenyl]-1H-pyrazol-4-yl}-(4-bromo-1H-indol-2-yl)-methanone

A mixture of{5-amino-1-[4-(2,3-difluoro-phenoxy)-2-methyl-phenyl]-1H-pyrazol-4-yl}-(1-benzenesulfonyl-4-bromo-1H-indol-2-yl)-methanone(275 mg, 0.414 mmol), Cs₂CO₃ (540 mg, 1.66 mmol), THF (10 mL) and MeOH(5 mL) was stirred at room temperature overnight. The solvent wasremoved under reduced pressure. The residue was purified bychromatography (silica gel, 40% EtOAc/hexanes) to give{5-amino-1-[4-(2,3-difluoro-phenoxy)-2-methyl-phenyl]-1H-pyrazol-4-yl}-(4-bromo-1H-indol-2-yl)-methanone(210 mg, 97%) as an off-white solid. ¹H NMR (400 MHz, DMSO-d₆) δ ppm12.10 (s, 1H), 8.28 (s, 1H), 7.51 (d, J=8.3 Hz, 1H), 7.25-7.42 (m, 4H),7.15-7.23 (m, 3H), 7.09 (t, J=7.5 Hz, 1H), 7.03 (d, J=8.5 Hz, 1H), 6.96(br, s., 2H), 2.09 (s, 3H). MS calcd. for C₂₅H₁₈BrF₂N₄O₂ [(M+H)₊] 523,obsd. 522.9.

Example I-2{5-Amino-1-[4-(2,3-difluoro-phenoxy)-2-methyl-phenyl]-1H-pyrazol-4-yl}-(4-phenyl-1H-indol-2-yl)-methanone

Step 1:{5-Amino-1-[4-(2,3-difluoro-phenoxy)-2-methyl-phenyl]-1H-pyrazol-4-yl}-(1-benzenesulfonyl-4-phenyl-1H-indol-2-yl)-methanone

A mixture of{5-amino-1-[4-(2,3-difluoro-phenoxy)-2-methyl-phenyl]-1H-pyrazol-4-yl}-(1-benzenesulfonyl-4-bromo-1H-indol-2-yl)-methanone(which may be prepared as described in Example I-1 Step 4; 30 mg, 0.045mmol), phenylboronic acid (11 mg, 0.09 mmol), Pd(PPh₃)₄ (5.2 mg, 0.004mmol), K₂CO₃ (25 mg, 0.18 mmol), water (0.5 mL), toluene (1 mL), andEtOH (1 mL) was heated at 90° C. overnight. Water (2 mL) was added, andthe mixture was extracted with EtOAc (2×5 mL). The organic layer wasevaporated. The residue was purified by chromatography (silica gel, 30%EtOAc/hexanes) to give{5-amino-1-[4-(2,3-difluoro-phenoxy)-2-methyl-phenyl]-1H-pyrazol-4-yl}-(1-benzenesulfonyl-4-phenyl-1H-indol-2-yl)-methanone(28 mg, 94%) as an oil.

Step 2:{5-Amino-1-[4-(2,3-difluoro-phenoxy)-2-methyl-phenyl]-1H-pyrazol-4-yl}-(4-phenyl-1H-indol-2-yl)-methanone

A mixture of{5-amino-1-[4-(2,3-difluoro-phenoxy)-2-methyl-phenyl]-1H-pyrazol-4-yl}-(1-benzenesulfonyl-4-phenyl-1H-indol-2-yl)-methanone(28 mg, 0.04 mmol), Cs₂CO₃ (41.4 mg, 0.13 mmol), THF (2 mL) and MeOH (1mL) was stirred at room temperature overnight. The solvent was removedunder reduced pressure. The residue was purified by chromatography(silica gel, 40% EtOAc/hexanes) to give{5-amino-1-[4-(2,3-difluoro-phenoxy)-2-methyl-phenyl]-1H-pyrazol-4-yl}-(4-phenyl-1H-indol-2-yl)-methanone(20 mg, 86%) as a light yellow oil. ¹H NMR (400 MHz, CDCl₃) δ ppm 9.65(s, 1H), 8.17 (s, 1H), 7.72. (dd, J=8.4, 1.4 Hz, 2H), 7.48-7.56 (m, 3H),7.37-7.46 (m, 3H), 7.31 (d, J=8.6 Hz, 1H), 7.26 (s, 1H), 7.23 (dd,J=6.8, 1.4 Hz, 1H), 7.07 (d, J=5.5 Hz, 2H), 6.99 (d, J=2.7 Hz, 1H), 6.92(dt, J=8.1, 2.6 Hz, 2H), 2.17 (s, 3H). MS calcd. for C₃₁H₂₃F₂N₄O₂[(M+H)⁺] 521, obsd. 521.1.

Example I-3{5-Amino-1-[4-(2,3-difluoro-phenoxy)-2-methyl-phenyl]-1H-pyrazol-4-yl}-(4-thiophen-3-yl-1H-indol-2-yl)-methanone

Step 1:{5-Amino-1-[4-(2,3-difluoro-phenoxy)-2-methyl-phenyl]-1H-pyrazol-4-yl}-(1-benzenesulfonyl-4-thiophen-3yl-1H-indol-2-yl)-methanone

A mixture of{5-amino-1-[4-(2,3-difluoro-phenoxy)-2-methyl-phenyl]-1H-pyrazol-4-yl}-(1-benzenesulfonyl-4-bromo-1H-indol-2-yl)-methanone(which may be prepared as described in Example I-1 Step 4; 95 mg, 0.14mmol), thiophene-3-boronic acid (37 mg, 0.29 mmol), Pd(PPh₃)₄ (16.5 mar,0.014 mmol), K₂CO₃ (79.2 mg, 0.57 mmol), water (1.5 mL), toluene (3 mL),and EtOH (3 mL) was heated at 90° C. overnight. Water (2 mL) was added,and the mixture was extracted with EtOAc (2×5 mL). The organic layer wasevaporated. The residue was purified by chromatography (silica gel, 40%EtOAc/hexanes) to give{5-amino-1-[4-(2,3-difluoro-phenoxy)-2-methyl-phenyl]-1H-pyrazol-4-yl}-(1-benzenesulfonyl-4-thiophen-3-yl-1H-indol-2-yl)-methanone(94 mg, 92%) as an oil.

Step 2:{5-Amino-1-[4-(2,3-difluoro-phenoxy)-2-methyl-phenyl]-1H-pyrazol-4-yl}-(4-thiophen-3-yl-1H-indol-2-yl)-methanone

A mixture of{5-amino-1-[4-(2,3-difluoro-phenoxy)-2-methyl-phenyl]-1H-pyrazol-4-yl}-(1-benzenesulfonyl-4-thiophen-3-yl-1H-indol-2-yl)-methanone(94 mg, 0.14 mmol), Cs₂CO₃ (137 mg, 0.42 mmol), THF (6 mL) and MeOH (3mL) was stirred at room temperature overnight. The solvent was removedunder reduced pressure. The residue was purified by chromatography(silica gel, 40% EtOAc/hexanes) to give{5-amino-1-[4-(2,3-difluoro-phenoxy)-2-methyl-phenyl]-1H-pyrazol-4-yl}-(4-thiophen-3-yl-1H-indol-2-yl)-methanone(70 mg, 88%) as an off-white solid. ¹H NMR (400 MHz, CDCl₃) δ ppm 9.40(s, 1H), 8.20 (s, 1H), 7.59-7.61 (m, 1H), 7.54 (dd, J=2.3, 0.8 Hz, 1H),7.48-7.53 (m, 2H), 7.40-7.44 (m, 1H), 7.35-7.40 (m, 1H), 7.27-7.34 (m,2H), 7.03-7.13 (m, 2H), 7.00 (d, J=2.7 Hz, 1H), 6.89-6.96 (m, 2H), 2.17(s, 3H). MS calcd. for C₂₉H₂₁F₂N₄O₂S [(M+H)⁺] 527, obsd. 527.

Example I-4{5-Amino-1-[4-(2,3-difluoro-phenoxy)-2-methyl-phenyl]-1H-pyrazol-4-yl}-(4-pyrazol-1-yl-1H-indol-2-yl)-methanone

A mixture of{5-amino-1-[4-(2,3-difluoro-phenoxy)-2-methyl-phenyl]-1H-pyrazol-4-yl}-(4-bromo-1H-indol-2-yl)-methanone(which may be prepared as described in Example I-1 Step 5; 100 mg, 0.15mmol), pyrazole (20.5 mg, 0.3 mmol), L-proline (7 mg, 0.06 mmol),copper(I) iodide (6 mg, 0.03 mmol), and K₂CO₃ (62.5 mg, 0.45 mmol) inDMSO (2 mL) was flushed with argon. The mixture was heated at 130° C.for 40 h and then purified by preparative HPLC to give{5-amino-1-[4-(2,3-difluoro-phenoxy)-2-methyl-phenyl]-1H-pyrazol-4-yl}-(4-pyrazol-1-yl-1H-indol-2-yl)-methanone(6 mg, 8%) as a yellow powder. ¹H NMR (400 MHz, CDCl₃) δ ppm 9.44 (br.s., 1H), 8.29 (s, 1H), 8.08 (dd, J=2.5, 0.5 Hz, 1H), 7.83-7.93 (m, 2H),7.71 (s, 1H), 7.39-7.48 (m, 2H), 7.31-7.37 (m, 2H), 7.05-7.17 (m, 2H),7.03 (d, J=2.8 Hz, 1H), 6.91-7.00 (m, 2H), 6.57-6.59 (m, 1H), 2.20 (s,3H), MS calcd. for C₂₈H₂₁F₂N₆O₂ [(M+H)⁺] 511, obsd. 511.

Example I-5{5-Amino-1-[4-(2,3-difluoro-phenoxy)-2-methyl-phenyl]-1H-pyrazol-4-yl}-(4-thiophen-2-yl-1H-indol-2-yl)-methanone

Step 1:{5-Amino-1-[4-(2,3-difluoro-phenoxy)-2-methyl-phenyl]-1H-pyrazol-4-yl}-(1-benzenesulfonyl-4-thiophen-2-yl-1H-indol-2-yl)-methanone

A mixture of{5-amino-1-[4-(2,3-difluoro-phenoxy)-2-methyl-phenyl]-1H-pyrazol-4-yl}-(1-benzenesulfonyl-4-bromo-1H-indol-2-yl)-methanone(which may be prepared as described in Example I-1 Step 4; 60 mg, 0.09mmol), 2-(tributylstannyl)thiophene (67.5 mg, 0.18 mmol), Pd(PPh₃)₄(10.4 mg, 0.009 mmol), and toluene (2 mL) was heated at 100° C.overnight. The solvent was evaporated. The residue was purified bychromatography (silica gel, 35% EtOAc/hexanes) to give{5-amino-1-[4-(2,3-difluoro-phenoxy)-2-methyl-phenyl]-1H-pyrazol-4-yl}-(1-benzenesulfonyl-4-thiophen-2-yl-1H-indol-2-yl)-methanone(46 mg, 76%) as an oil.

Step 2:{5-Amino-1-[4-(2,3-difluoro-phenoxy)-2-methyl-phenyl]-1H-pyrazol-4-yl}-(4-thiophen-2-yl-1H-indol-2-yl)-methanone

A mixture of{5-amino-1-[4-(2,3-difluoro-phenoxy)-2-methyl-phenyl]-1H-pyrazol-4-yl}-(1-benzenesulfonyl-4-thiophen-2-yl-1H-indol-2-yl)-methanone(46 mg, 0.07 mmol), Cs₂CO₃ (67.4 mg, 0.21 mmol), THF (2 mL) and MeOH (1mL) was stirred at room temperature overnight. The solvent was removedunder reduced pressure. The residue was purified by chromatography(silica gel, 40% EtOAc/hexanes) to give{5-amino-1-[4-(2,3-difluoro-phenoxy)-2-methyl-phenyl]-1H-pyrazol-4-yl}-(4-thiophen-2-yl-1H-indol-2-yl)-methanone(32 mg, 88%) as an off-white solid. ¹H NMR (400 MHz, CDCl₃) δ ppm 9.41(br. s., 1H), 8.24 (s, 1H), 7.73 (dd, J=2.0, 0.8 Hz, 1H), 7.49 (dd,J=3.5, 1.2 Hz, 1H), 7.40-7.44 (m, 2H), 7.36-7.39 (m, 2H), 7.30-7.36 (m,2H), 7.20 (dd, J=5.1, 3.5 Hz, 1H), 7.02-7.13 (m, 2H), 7.00 (d, J=2.7 Hz,1H), 6.88-6.96 (m, 2H), 2.18 (s, 3H). MS calcd. for C₂₉H₂₁F₂N₄O₂S[(M+H)⁺]527, obsd. 526.9.

Example I-6{5-Amino-1-[4-(2,3-difluoro-phenoxy)-2-methyl-phenyl]-1H-pyrazol-4-yl}-[4-(morpholine-4-carbonyl)-1H-indol-2-yl]-methanonetrifluoroacetate salt

Carbon monoxide gas was bubbled through a mixture of{5-amino-1-[4-(2,3-difluoro-phenoxy)-2-methyl-phenyl]-1H-pyrazol-4-yl}-(4-bromo-1H-indol-2-yl)-methanone(which may be prepared as described in Example I-1 Step 5; 94 mg, 0.18mmol), morpholine (313 mg, 3.6 mmol), Pd(PPh₃)₄ (62.3 mg, 0.054 mmol),and THF (10 mL) in a sealable tube for 5 min. The tube was sealed andheated at 90° C. for 2 h. The solvent was evaporated and the residue waspurified by preparative HPLC to give{5-amino-1-[4-(2,3-difluoro-phenoxy)-2-methyl-phenyl]-1H-pyrazol-4-yl}-[4-(morpholine-4-carbonyl)-1H-indol-2-yl]-methanonetrifluoroacetate salt (24 mg, 24%) as a light yellow powder. ¹H NMR (400MHz, CDCl₃) δ ppm 9.52 (s, 1H), 8.23 (s, 1H), 7.51 (d, J=8.2 Hz, 1H),7.30-7.38 (m, 3H), 7.18 (dd, J=7.2, 1.0 Hz, 1H), 7.02-7.15 (m, 2H), 7.00(d, J=2.7 Hz, 1H), 6.88-6.96 (m, 2H), 3.41-4.00 (m, 8H), 2.17 (s, 3H).MS calcd. for C₃₀H₂₆F₂N₅O₄ [(M+H)⁺]558, obsd. 558.

Example I-7{5-Amino-1-[4-(2,3-difluoro-phenoxy)-2-methyl-phenyl]-1H-pyrazol-4-yl}-[4-(1-methyl-1H-pyrazol-3-yl)-1H-indol-2-yl]-methanone

Step 1:{5-Amino-1-[4-(2,3-difluoro-phenoxy)-2-methyl-phenyl]-1H-pyrazol-4-yl}-[1-benzenesulfonyl-4-(1-methyl-1H-pyrazol-3-yl)-1H-indol-2-yl]-methanone

A mixture of{5-amino-1-[4-(2,3-difluoro-phenoxy)-2-methyl-phenyl]-1H-pyrazol-4-yl}-(1-benzenesulfonyl-4-bromo-1H-indol-2-yl)-methanone(which may be prepared as described in Example I-1 Step 4; 65 mg, 0.098mmol), 1-methyl-1H-pyrazole-4-boronic acid hydrochloride (32 mg, 0.196mmol), Pd(PPh₃)₄ (11.3 mg, 0.01 mmol), K₂CO₃ (54.2 mg, 0.39 mmol), water(1 mL), toluene (2 mL), and EtOH (2 mL) was heated at 90° C. overnight.Water (2 mL) was added, and the mixture was extracted with EtOAc (2×5mL). The organic layer was evaporated. The residue was purified bychromatography (silica gel, 40% EtOAc/hexanes) to give{5-amino-1-[4-(2,3-difluoro-phenoxy)-2-methyl-phenyl]-1H-pyrazol-4-yl}-[1-benzenesulfonyl-4-(1-methyl-1H-pyrazol-3-yl)-1H-indol-2-yl]-methanone(31 mg, 48%) as an oil.

Step 2:{5-Amino-1-[4-(2,3-difluoro-phenoxy)-2-methyl-phenyl]-1H-pyrazol-4-yl}-[4-(1-methyl-1H-pyrazol-3-yl)-1H-indol-2yl]-methanone

A mixture of{5-amino-1-[4-(2,3-difluoro-phenoxy)-2-methyl-phenyl]-1H-pyrazol-4-yl}-[1-benzenesulfonyl-4-(1-methyl-1H-pyrazol-3-yl)-1H-indol-2-yl]-methanone(31 mg, 0.046 mmol), Cs₂CO₃ (60.8 mg, 0.19 mmol), THF (1.5 mL) and MeOH(0.75 mL) was stirred at room temperature overnight. The solvent wasremoved under reduced pressure. The residue was purified bychromatography (silica gel, 90% EtOAc/hexanes) to give{5-amino-1-[4-(2,3-difluoro-phenoxy)-2-methyl-phenyl]-1H-pyrazol-4-yl}-[4-(1-methyl-1H-pyrazol-3-yl)-1H-indol-2-yl]-methanone(21 mg, 86%) as an off-white solid. ¹H NMR (400 MHz, CDCl₃) δ ppm 9.29(s, 1H), 8.21 (s, 1H), 7.93 (s, 1H), 7.78 (s, 1H), 7.47-7.50 (m, 1H),7.30-7.37 (m, 3H), 7.22 (dd, J=6.4, 1.8 Hz, 1H), 7.02-7.14 (m, 2H), 7.00(d, J=2.7 Hz, 1H), 6.86-6.96 (m, 2H), 4.05 (s, 3H), 2.18 (s, 3H). MScalcd, for C₂₉H₂₃F₂N₆O₂ [(M+H)⁺] 525, obsd. 525.

Example I-8{5-Amino-1-[4-(2,3-difluoro-phenoxy)-2-methyl-phenyl]-1H-pyrazol-4-yl}-(4-pyridin-2-yl-1H-indol-2-yl)-methanone

Step 1:{5-Amino-1-[4-(2,3-difluoro-phenoxy)-2-methyl-phenyl]-1H-pyrazol-4-yl}-(1-benzenesulfonyl-4-pyridin-2-yl-1H-indol-2-yl)-methanone

A mixture of{5-amino-1-[4-(2,3-difluoro-phenoxy)-2-methyl-phenyl]-1H-pyrazol-4-yl}-(1-benzenesulfonyl-4-bromo-1H-indol-2-yl)-methanone(which may be prepared as described in Example I-1 Step 4; 64 mg, 0.097mmol), 2-(tributylstannyl)pyridine (71 mg, 019 mmol), Pd(PPh₃)₄ (11.1mg, 0.01 mmol), and toluene (2 mL) was heated at 100° C. overnight. Thesolvent was evaporated. The residue was purified by chromatography(silica gel, 45% EtOAc/hexanes) to give{5-amino-1-[4-(2,3-difluoro-phenoxy)-2-methyl-phenyl]-1H-pyrazol-4-yl}-(1-benzenesulfonyl-4-pyridin-2-yl-1H-indol-2-yl)-methanone(27 mg, 42%) as an oil.

Step 2:{5-Amino-1-[4-(2,3-difluoro-phenoxy)-2-methyl-phenyl]-1H-pyrazol-4-yl}-(4-pyridin-2-yl-1H-indol-2-yl)-methanone

A mixture of{5-amino-1-[4-(2,3-difluoro-phenoxy)-2-methyl-phenyl]-1H-pyrazol-4-yl}-(1-benzenesulfonyl-4-pyridin-2-yl-1H-indol-2-yl)-methanone(27 mg, 0.041 mmol), Cs₂CO₃ (53.2 mg, 0.16 mmol), THF (2 mL) and MeOH (1mL) was stirred at room temperature overnight. The solvent was removedunder reduced pressure. The residue was purified by chromatography(silica gel, 50% EtOAc/hexanes) to give{5-amino-1-[4-(2,3-difluoro-phenoxy)-2-methyl-phenyl]-1H-pyrazol-4-yl}-(4-pyridin-2-yl-1H-indol-2-yl)-methanone(19 mg, 79%; purity 88%) as an off-white solid. ¹H NMR (400 MHz, CDCl₃)δ ppm 9.37 (br. s., 1H), 8.87 (d, J=4.8 Hz, 1H), 8.29 (s, 1H), 7.91 (d,J=18.1 Hz, 3H), 7.44-7.65 (m, 3H), 7.35 (d, J=8.5 Hz, 2H), 7.04-7.18 (m,2H), 7.01-7.04 (m, 1H), 6.88-6.99 (m, 2H), 2.20 (s, 3H). MS calcd. forC₃₀H₂₂F₂N₅O₂ [(M+H)⁺] 522, obsd. 522.

Example I-9{5-Amino-1-[4-(2,3-difluoro-phenoxy)-2-methyl-phenyl]-1H-pyrazol-4-yl}-(4-benzyl-1H-indol-2-yl)-methanone

Step 1:{5-Amino-1-[4-(2,3-difluoro-phenoxy)-2-methyl-phenyl]-1H-pyrazol-4-yl}-(1-benzenesulfonyl-4-benzyl-1H-indol-2-yl)-methanone

A mixture of{5-amino-1-[4-(2,3-difluoro-phenoxy)-2-methyl-phenyl]-1H-pyrazol-4-yl}-(1-benzenesulfonyl-4-bromo-1H-indol-2-yl)-methanone(which may be prepared as described in Example I-1 Step 4; 65 mg, 0.098mmol), Pd(OAc)₂ (2.8 mg, 0.012 mmol),2-dicyclohexylphosphino-2′,6′-dimethoxybiphenyl (Aldrich; 9 mg, 0.022mmol), and THF (2 mL) was flushed with argon for 3 min and then stirredat room temperature for 5 min. Benzylzinc(II) bromide (Aldrich; 0.5 M inTHF; 0.3 mL; 0.15 mmol) was added and the mixture was heated at 75° C.for 2 h. The solvent was evaporated. The residue was purified bychromatography (silica gel, 35% EtOAc/hexanes) to give{5-amino-1-[4-(2,3-difluoro-phenoxy)-2-methyl-phenyl]-1H-pyrazol-4-yl}-(1-benzenesulfonyl-4-benzyl-1H-indol-2-yl)-methanone(66 mg, 82%) as an oil.

Step 2:{5-Amino-1-[4-(2,3-difluoro-phenoxy)-2-methyl-phenyl]-1H-pyrazol-4-yl}-(4-benzyl-1H-Indol-2-yl)-methanone

A mixture of{5-amino-1-[4-(2,3-difluoro-phenoxy)-2-methyl-phenyl]-1H-pyrazol-4-yl}-(1-benzenesulfonyl-4-benzyl-1H-indol-2-yl)-methanone(63 mg, 0.093 mmol), Cs₂CO₃ (122 mg, 0.37 mmol), THF (5 mL) and MeOH(2.5 mL) was stirred at room temperature overnight. The solvent wasremoved under reduced pressure. The residue was purified bychromatography (silica gel, 40% EtOAc/hexanes) to give{5-amino-1-[4-(2,3-difluoro-phenoxy)-2-methyl-phenyl]-1H-pyrazol-4-yl}-(4-benzyl-1H-indol-2-yl)-methanone(42 mg, 84%) as a yellow oil. ¹H NMR (400 MHz, CDCl₃) δ ppm 9.26 (br.s., 1H), 8.06 (s, 1H), 7.30-7.42 (m, 8H), 7.25 (dd, J=2.1, 0.9 Hz, 2H),7.05-7.17 (m, 2H), 6.91-7.04 (m, 4H), 4.36 (s, 2H), 2.19 (s, 3H). MScalcd. for C₃₂H₂₅F₂N₄O₂ [(M+H)⁺] 535, obsd. 536.

Example I-10{5-Amino-1-[4-(2,3-difluoro-phenoxy)-2-methyl-phenyl]-1H-pyrazol-4-yl}-[4-(1H-pyrazol-4-yl)-1H-indol-2-yl]-methanone

A mixture of{5-amino-1-[4-(2,3-difluoro-phenoxy)-2-methyl-phenyl]-1H-pyrazol-4-yl}-(4-bromo-1H-indol-2-yl)-methanone(which may be prepared as described in Example I-1 Step 5; 55 mg, 0.105mmol), tert-butoxycarbonyl-1H-pyrazole-4-boronic acid (44.6 mg, 0.21mmol), Pd(PPh₃)₄ (12.1 mg, 0.011 mmol), K₂CO₃ (58.1 mg, 0.42 mmol),water (0.75 mL), toluene (1.5 mL), and EtOH (1.5 mL) was heated at 90°C. overnight. Water (2 mL) was added, and the mixture was extracted withEtOAc (2×5 mL). The organic layer was evaporated. The residue waspurified by chromatography (silica gel, 60% EtOAc/hexanes) to give{5-amino-1-[4-(2,3-difluoro-phenoxy)-2-methyl-phenyl]-1H-pyrazol-4-yl}-[4-(1H-pyrazol-4-yl)-1H-indol-2-yl]-methanone(30 mg, 56%) as an oil, ¹H NMR (400 MHz, DMSO-d₆) δ ppm 13.04 (br. s.,1H), 11.78 (s, 1H), 8.39-8.47 (m, 2H), 8.07 (s, 1H), 7.23-7.49 (m, 7H),7.17 (d, J=2.8 Hz, 1H), 6.99-7.13 (m, 2H), 6.90 (s, 2H), 2.10 (s, 3H).MS calcd. for C₂₈H₂₁F₂N₆O₂ [(M+H)⁺] 511, obsd. 511.

Example I-113-(2-{5-Amino-1-[4-(2,3-difluoro-phenoxy)-2-methyl-phenyl]-1H-pyrazole-4-carbonyl}-1H-indol-4-yl)-benzonitrile

A mixture of{5-amino-1-[4-(2,3-difluoro-phenoxy)-2-methyl-phenyl]-1H-pyrazol-4-yl}-(4-bromo-1H-indol-2-yl)-methanone(which may be prepared as described in Example I-1 Step 5; 55 mg 0.105mmol), 3-cyanophenylboronic acid (30.9 mg, 0.21 mmol), Pd(PPh₃)₄ (12.1mg, 0.011 mmol), K₂CO₃ (58.1 mg, 0.42 mmol), water (0.75 mL), toluene(1.5 mL), and EtOH (1.5 mL) was heated at 90° C. overnight. Water (2 mL)was added, and the mixture was extracted with EtOAc (2×5 mL). Theorganic layer was evaporated. The residue was purified by chromatography(silica gel, 35% EtOAc/hexanes) to give3-(2-{5-amino-1-[4-(2,3-difluoro-phenoxy)-2-methyl-phenyl]-1H-pyrazole-4-carbonyl}-1H-indol-4-yl)-benzonitrile(38 mg, 66%) as an oil. ¹H NMR (400 MHz, CDCl₃) δ ppm 9.43 (s, 1H), 8.15(s, 1H), 7.96-8.01 (m, 2H), 7.73-7.78 (m, 1H), 7.64-7.70 (m, 1H),7.53-7.57 (m, 1H), 7.42-7.49 (m, 1H), 7.32-7.38 (m, 2H), 7.23 (dd,J=7.2, 0.9 Hz, 1H), 7.05-7.17 (m, 2H), 7.02 (d, J=3.0 Hz, 1H), 6.91-6.99(m, 2H), 2.20 (s, 3H). MS calcd. for C₃₂H₂₂F₂N₅O₂ [(M+H)⁺] 546, obsd.546.

Example I-12{5-Amino-1-[4-(2,3-difluoro-phenoxy)-2-methyl-phenyl]-1H-pyrazol-4-yl}-[4-(3-chloro-phenyl)-1H-indol-2-yl]-methanone

A mixture of{5-amino-1-[4-(2,3-difluoro-phenoxy)-2-methyl-phenyl]-1H-pyrazol-4-yl}-(4-bromo-1H-indol-2-yl)-methanone(which may be prepared as described in Example I-1 Step 5; 55 mg, 0.105mmol), 3-chlorophenylboronic acid (32.9 mg, 0.21 mmol), Pd(PPh₃)₄ (12.1mg, 0.011 mmol), K₂CO₃ (58.1 mg, 0.42 mmol), water (0.75 mL), toluene(1.5 mL), and EtOH (1.5 mL) was heated at 90° C. overnight. Water (2 mL)was added, and the mixture was extracted with EtOAc (2×5 mL). Theorganic layer was evaporated. The residue was purified by chromatography(silica gel, 35% EtOAc/hexanes) to give{5-amino-1-[4-(2,3-difluoro-phenoxy)-2-methyl-phenyl]-1H-pyrazol-4-yl}-[4-(3-chloro-phenyl)-1H-indol-2-yl]-methanone(54 mg, 87%) as an oil. ¹H NMR (400 MHz, CDCl₃) δ ppm 9.41 (br. s., 1H),8.17 (s, 1H), 7.71 (t, J=1.8 Hz, 1H), 7.63 (dt, J=7.5, 1.4 Hz, 1H),7.48-7.53 (m, 2H), 7.41-7.47 (m, 3H), 7.34 (d, J=8.5 Hz, 1H), 7.24 (dd,J=7.3, 1.0 Hz, 1H), 7.05-7.16 (m, 2H), 7.02 (d, J=2.5 Hz, 1H), 6.90-6.99(m, 2H), 2.20 (s, 3H). MS calcd. for C₃₁H₂₂ClF₂N₄O₂ [(M+H)⁺] 555, obsd.555.

Example I-133-(2-{5-Amino-1-[4-(2,3-difluoro-phenoxy)-2-methyl-phenyl]-1H-pyrazole-4-carbonyl}-1H-indol-4-ylmethyl)-benzonitrile

Step 1:3-(2-{5-Amino-1-[4-(2,3-difluoro-phenoxy)-2-methyl-phenyl]-1H-pyrazole-4-carbonyl}-1-benzenesulfonyl-1H-indol-4-ylmethyl)-benzonitrile

A mixture of{5-amino-1-[4-(2,3-difluoro-phenoxy)-2-methyl-phenyl]-1H-pyrazol-4-yl}-(1-benzenesulfonyl-4-bromo-1H-indol-2-yl)-methanone(which may be prepared as described in Example I-1 Step 4; 65 mg, 0.098mmol), Pd(OAc)₂ (2.8 mg, 0.012 mmol),2-dicyclohexylphosphino-2′,6′-dimethoxybiphenyl (Aldrich; 9 mg, 0.022mmol), and THF (2 mL) was flushed with argon for 3 min and then stirredat room temperature for 5 min. (3-Cyano-benzyl)zinc(II) bromide(Aldrich; 0.5 M in THF; 0.3 mL; 0.15 mmol) was added and the mixture washeated at 75° C. for 2 h. Water (3 mL) was added and the mixture wasextracted with EtOAc (2×5 mL). The combined organic layers wereevaporated. The residue was purified by chromatography (silica gel, 35%EtOAc/hexanes) to give3-(2-{5-amino-1-[4-(2,3-difluoro-phenoxy)-2-methyl-phenyl]-1H-pyrazole-4-carbonyl}-1-benzenesulfonyl-1H-indol-4-ylmethyl)-benzonitrile(64 mg, 93%) as an oil.

Step 2:3-(2-{5-Amino-1-[4-(2,3-difluoro-phenoxy)-2-methyl-phenyl]-1H-pyrazole-4-carbonyl}-1H-indol-4-ylmethyl)-benzonitrile

A mixture of3-(2-{5-amino-1-[4-(2,3-difluoro-phenoxy)-2-methyl-phenyl]-1H-pyrazole-4-carbonyl}-1-benzenesulfonyl-1H-indol-4-ylmethyl)-benzonitrile(64 mg, 0.092 mmol), Cs₂CO₃ (119 mg, 0.37 mmol), THF (5 mL) and MeOH(2.5 mL) was stirred at room temperature overnight. The solvent wasremoved under reduced pressure. The residue was purified bychromatography (silica gel, 40% EtOAc/hexanes) to give3-(2-{5-amino-1-[4-(2,3-difluoro-phenoxy)-2-methyl-phenyl]-1H-pyrazole-4-carbonyl}-1H-indol-4-ylmethyl)-benzonitrile(40 mg, 78%) as a yellow oil. ¹H NMR (400 MHz, CDCl₃) δ ppm 9.30 (br.s., 1H), 8.07 (s, 1H), 7.50-7.60 (m, 3H), 7.43 (dd, J=11.2, 8.2 Hz, 2H),7.30-7.36 (m, 2H), 7.20 (s, 1H), 7.10 (ddd, J=15.2, 8.7, 5.9 Hz, 2H),7.02 (d, J=2.5 Hz, 1H), 6.90-6.98 (m, 3H), 4.39 (s, 2H), 2.19 (s, 3H).MS calcd. for C₃₃H₂₄F₂N₅O₂ [(M+H)⁺] 560, obsd. 560.1.

Example I-143-(4-{5-Amino-4-[4-(1H-pyrazol-4-yl)-1H-indole-2-carbonyl]-pyrazol-1-yl}-3-methyl-phenoxy)-benzonitrile

Step 1:3-(4-[5-Amino-4-(1-benzenesulfonyl-4-bromo-1H-indole-2-carbonyl)-pyrazol-1-yl]-3-methyl-phenoxyl-benzonitrile

A mixture of 3-(4-hydrazino-3-methyl-phenoxy)-benzonitrile (which may beprepared as described for Intermediate 2; 789 mg, 3.3 mmol),(E)-2-(1-benzenesulfonyl-4-bromo-1H-indole-2-carbonyl)-3-dimethylamino-acrylonitrile(which may be prepared as described in Example I-1 Step 3; 510 mg, 1.11mmol) and EtOH (30 mL) was heated at reflux overnight. The solvent wasremoved under reduced pressure. The residue was purified bychromatography (silica gel, 30% EtOAc/hexanes) to give3-{4-[5-amino-4-(1-benzenesulfonyl-4-bromo-1H-indole-2-carbonyl)-pyrazol-1-yl]-3-methyl-phenoxy}-benzonitrile(598 mg, 82%) as a foam.

Step 2:3-{4-[5-Amino-4-(4-bromo-1H-indole-2-carbonyl)-pyrazol-1-yl]-3-methyl-phenoxy}-benzonitrile

A mixture of3-{4-[5-amino-4-(1-benzenesulfonyl-4-bromo-1H-indole-2-carbonyl)-pyrazol-1-yl]-3-methyl-phenoxy}-benzonitrile(450 mg, 0.69 mmol), Cs₂CO₃ (899 mg, 2.76 mmol), THF (15 mL) and MeOH(7.5 mL) was stirred at room temperature overnight. The solvent wasremoved under reduced pressure. The residue was purified bychromatography (silica gel, 40% EtOAc/hexanes) to give3-{4-[5-amino-4-(4-bromo-1H-indole-2-carbonyl)-pyrazol-1-yl]-3-methyl-phenoxy}-benzonitrile.

Step 3:3-(4-{5-Amino-4-[4-(1H-pyrazol-4-yl)-1H-indole-2-carbonyl]-pyrazol-1-yl}-3-methyl-phenoxy)-benzonitrile

A mixture of3-{4-[5-amino-4-(4-bromo-1H-indole-2-carbonyl)-pyrazol-1-yl]-3-methyl-phenoxy}-benzonitrile(55 mg, 0.107 mmol), 1-tert-butoxycarbonyl-1H-pyrazole-4-boronic acid(Combi-Blocks Inc., 7949 Silverton Avenue, Suite 915, San Diego, Calif.92126, USA; 45.5 mg, 0.22 mmol), Pd(PPh₃)₄ (12.4 mg, 0.011 mmol), K₂CO₃(59.3 mg, 0.43 mmol), water (0.75 mL), toluene (1.5 mL), and EtOH (1.5mL) was heated at 90° C. overnight. Water (2 mL) was added, and themixture was extracted with EtOAc (2×5 mL). The organic layer wasevaporated. The residue was purified by chromatography (silica gel, 80%EtOAc/hexanes) to give3-(4-{5-amino-4-[4-(1H-pyrazol-4-yl)-1H-indole-2-carbonyl]-pyrazol-1-yl}-3-methyl-phenoxy)-benzonitrile(30 mg, 56%) as an oil. ¹H NMR (400 MHz, DMSO-d₆) δ ppm 13.04 (br. s.,1H), 11.78 (s, 1H), 8.38-8.49 (m, 2H), 8.08 (br. s., 1H), 7.56-7.71 (m,3H), 7.35-7.52 (m, 4H), 7.19-7.30 (m, 3H), 7.08 (dd, J=8.5, 2.8 Hz, 1H),6.92 (s, 2H), 2.11 (s, 3H). MS calcd. for C₂₉H₂₂N₇O₂ [(M+H)⁺] 500, obsd.500.

Example I-153-(4-{5-Amino-4-[4-(morpholine-4-carbonyl)-1H-indole-2-carbonyl]-pyrazol-1-yl}-3-methyl-phenoxy)-benzonitrile

Carbon monoxide gas was bubbled through a mixture of3-{4-[5-amino-4-(4-bromo-1H-indole-2-carbonyl)-pyrazol-1-yl]-3-methyl-phenoxy}-benzonitrile(which may be prepared as described in Example I-14 Step 2; 53 mg, 0.10mmol), morpholine (180 mg, 2.1 mmol), Pd(PPh₃)₄ (35.9 mg. 0.031 mmol),and THF (10 mL) in a sealable tube for 5 min. The tube was sealed andheated at 90° C. overnight. The solvent was evaporated and the residuewas purified by preparative HPLC to give3-(4-{5-amino-4-[4-(morpholine-4-carbonyl)-1H-indole-2-carbonyl]-pyrazol-1-yl}-3-methyl-phenoxy)-benzonitrile(21 mg, 37%) as a light yellow powder. MS calcd. for C₃₁H₂₇N₆O₄ [(M+H)⁺]547, obsd. 546.9.

Example I-163-(4-{5-Amino-4-[4-(4-methyl-piperazine-4-carbonyl)-1H-indole-2-carbonyl]-pyrazol-1-yl}-3-methyl-phenoxy)-benzonitrile

Carbon monoxide gas was bubbled through a mixture of3-{4-[5-amino-4-(4-bromo-1H-indole-2-carbonyl)-pyrazol-1-yl]-3-methyl-phenoxy}-benzonitrile(which may be prepared as described in Example I-14 Step 2; 53 mg, 0.10mmol), 1-methylpiperazine (155 mg, 1.55 mmol), Pd(PPh₃)₄ (35.9 mg, 0.031mmol), and THF (10 mL) in a sealable tube for 5 min. The tube was sealedand heated at 90° C. overnight. The solvent was evaporated and theresidue was purified by preparative HPLC to give3-(4-{5-amino-4-[4-(4-methyl-piperazine-1-carbonyl)-1H-indole-2-carbonyl]-pyrazol-1-yl}-3-methyl-phenoxy)-benzonitrile(32 mg, 55%) as an off-white foam. MS calcd. for C₃₂H₃₀N₇O₃ [(M+H)⁺]560, obsd. 560.

Example I-173-(4-{5-Amino-4-[4-(3-methoxy-benzyl)-1H-indole-2-carbonyl]-pyrazol-1-yl}-3-methyl-phenoxy)-benzonitrile

Step 1:3-(4-{5-Amino-4-[1-benzenesulfonyl-4-(3-methoxy-benzyl)-1H-indole-2-carbonyl]-pyrazol-1-yl}-3-methyl-phenoxy)-benzonitrile

A mixture of3-{4-[5-amino-4-(1-benzenesulfonyl-4-bromo-1H-indole-2-carbonyl)-pyrazol-1-yl]-3-methyl-phenoxy}-benzonitrile(which may be prepared as described in Example I-14 Step 1; 65 mg, 0.10mmol), Pd(OAc)₂ (2.8 mg, 0.012 mmol),2-dicyclohexylphosphino-2′,6′-dimethoxybiphenyl (Aldrich; 9 mg, 0.022mmol), and THF (2 mL) was flushed with argon for 5 min and then stirredat room temperature for 5 min (3-Methoxy-benzyl)zinc(II) bromide(Aldrich; 0.5 M in THF; 0.3 mL; 0.15 mmol) was added and the mixture washeated at 75° C. for 2 h. Water (3 mL) was added and the mixture wasextracted with EtOAc (2×5 mL). The combined organic layers wereevaporated. The residue was purified by chromatography (silica gel, 40%EtOAc/hexanes) to give3-(4-{5-amino-4-[1-benzenesulfonyl-4-(3-methoxy-benzyl)-1H-indole-2-carbonyl]-pyrazol-1-yl}-3-methyl-phenoxy)-benzonitrile(62 mg, 90%) as an oil.

Step 2:3-(4-{5-Amino-4-[4-(3-methoxy-benzyl)-1H-indole-2-carbonyl]-pyrazol-1-yl}-3-methyl-phenoxy)-benzonitrile

A mixture of3-(4-{5-amino-4-[1-benzenesulfonyl-4-(3-methoxy-benzyl)-1H-indole-2-carbonyl]-pyrazol-1-yl}-3-methyl-phenoxy)-benzonitrile(62 mg, 0.089 mmol), Cs₂CO₃ (116 mg, 0.36 mmol), THF (5 mL) and MeOH(2.5 mL) was stirred at room temperature overnight. The solvent wasremoved under reduced pressure. The residue was purified bychromatography (silica gel, 40% EtOAc/hexanes) to give3-(4-{5-amino-4-[4-(3-methoxy-benzyl)-1H-indole-2-carbonyl]-pyrazol-1-yl}-3-methyl-phenoxy)-benzonitrile(41 mg, 83%) as a yellow foam. ¹H NMR (400 MHz, CDCl₃) δ ppm 9.28 (s,1H), 8.11 (s, 1H), 7.44-7.56 (m, 2H), 7.37-7.42 (m, 2H), 7.30-7.37 (m,4H), 7.25 (d, J=8.0 Hz, 1H), 6.91-7.08 (m, 5H), 6.77-6.87 (m, 2H), 4.34(s, 2H), 3.78 (s, 3H), 2.22 (s, 3H). MS calcd. for C₃₄H₂₈N₅O₃ [(M+H)⁺]554, obsd. 554.1.

Example I-182-{5-Amino-1-[4-(2,3-difluoro-phenoxy)-2-methyl-phenyl]-1H-pyrazole-4-carbonyl}-1H-indole-4-carboxylicacid methyl ester

Carbon monoxide gas was bubbled through a mixture of{5-amino-1-[4-(2,3-difluoro-phenoxy)-2-methyl-phenyl]-1H-pyrazol-4-yl}-(4-bromo-1H-indol-2-yl)-methanonewhich may be prepared as described in Example I-1 Step 5; 200 mg, 0.38mmol), MeOH (2.5 mL, 62 mmol), Pd(PPh₃)₄ (132 mg, 0.115 mmol), and THF(20 mL) in a sealable tube for 5 min. The tube was sealed and heated at90° C. overnight. The solvent was evaporated and the residue waspurified by chromatography (silica gel, 45% EtOAc/hexanes) to give2-{5-amino-1-[4-(2,3-difluoro-phenoxy)-2-methyl-phenyl]-1H-pyrazole-4-carbonyl}-1H-indole-4-carboxylicacid methyl ester (101 mg, 45%; 85% purity) as an oil. ¹H NMR (400 MHz,CDCl₃) δ ppm 9.52 (br. s., 1H), 8.26 (s, 1H), 7.87-7.95 (m, 2H), 7.60(d, J=8.3 Hz, 2H), 7.43 (d, J=5.5 Hz, 2H), 7.23-7.35 (m, 3H), 7.19 (s,2H), 7.11 (d, J=7.5 Hz, 2H), 6.96-7.05 (m, 3H), 6.94 (d, J=2.8 Hz, 1H),6.79-6.90 (m, 2H), 3.96 (s, 3H), 2.11 (s, 3H), MS calcd. forC₂₇H₂₁F₂N₄O₄ [(M+H)⁺] 503, obsd. 502.9.

Example I-19{5-Amino-1-[4-(2,3-difluoro-phenoxy)-2-methyl-phenyl]-1H-pyrazol-4-yl}-(4-morpholin-4-ylmethyl-1H-indol-2-yl)-methanone

Step 1: (1-Benzenesulfonyl-1H-indol-4-yl)-morpholin-4-yl-methanone

A sealed tube containing 1-benzenesulfonyl-4-promo-1H-indole (which maybe prepared as described in Bell, I. M. et al. WO 2007061694 Page 103;5.18 g, 59.5 mmol), Pd(PPh₃)₄ (1.37 g, 1.19 mmol), and THF (125 mL) wascharged with 40 psi of carbon monoxide. The mixture was heated at 95° C.overnight and then the solvent was evaporated. The residue was purifiedby chromatography (silica gel, 80% EtOAc/hexanes) to give(1-benzenesulfonyl-1H-indol-4-yl)-morpholin-4-yl-methanone (2.67 g, 61%)as a foam.

Step 2: (1-Benzenesulfonyl-1H-indol-4-yl)-methanol

A solution of(1-benzenesulfonyl-1H-1-indol-4-yl)-morpholin-4-yl-methanone (2.07 g,5.59 mmol) in THF (50 mL) was cooled to −20° C. and LiAlH₄ (2 M in THF;6.2 mL, 12.4 mmol) was added dropwise. The mixture was stirred for 30min and MeOH (20 ML) was added. NaBH₄ (634 mg, 16.8 mmol) was added andthe mixture was stirred for 30 min. EtOAc (300 mL) was added and themixture was washed with brine. The organic layer was dried (Na₂SO₄),filtered, and evaporated to give(1-benzenesulfonyl-1H-indol-4-yl)-methanol (1.41 g, 88%) as an oil whichwas used directly in the next step without further purification.

Step 3: 1-Benzenesulfonyl-4-chloromethyl-1H-indole

DIPEA (3.17 g, 24.5 mmol) and methanesulfonyl chloride (1.69 g, 14.7mmol) were added to a solution of(1-benzenesulfonyl-1H-indol-4-yl)-methanol (1.41 g, 4.91 mmol) in THF(40 mL). The mixture was stirred at room temperature overnight. Thesolvent was evaporated under reduced pressure. The residue was purifiedby chromatography (silica gel, 20% EtOAc/hexanes) to give1-benzenesulfonyl-4-chloromethyl-1H-indole (1.4 g, 93%) as an oil.

Step 4: 1-Benzenesulfonyl-4-morpholin-4-ylmethyl-1H-indole

A mixture of 1-benzenesulfonyl-4-chloromethyl-1H-indole (1.4 g, 4.58mmol), morpholine (1.2 mL, 13.7 mmol), K₂CO₃ (3.16 g, 22.9 mmol) andCH₃CN (60 mL) was heated at 65° C. overnight. EtOAc (250 mL) was addedand the mixture was washed with water and brine, dried (Na₂SO₄),filtered, and evaporated. The residue was purified by chromatography(silica gel, 60% EtOAc/hexanes) to give1-benzenesulfonyl-4-morpholin-4-ylmethyl-1H-indole (1.45 g, 89%) as anoil.

Step 5: 1-Benzenesulfonyl-4-morpholin-4-ylmethyl-1H-indole-2-carboxylicadd

A solution of 1-benzenesulfonyl-4-morpholin-4-ylmethyl-1H-indole (600mg, 1.68 mmol) in THF (12 mL) was cooled to −78° C. LDA (2M in THF; 1.7mL, 3.4 mmol) was added and the mixture was stirred for 1 h. Excesssolid CO₂ was added. After 30 min at −78° C., the reaction mixture waswarmed to room temperature and 2 N HCl was added. The mixture wasextracted with CH₂Cl₂ (3×150 mL). The organic layer was dried (Na₂SO₄),filtered, and evaporated to give1-benzenesulfonyl-4-morpholin-4-ylmethyl-1H-indole-2-carboxylic acid(670 mg, 99%) as a foam which was used directly in the next step withoutfurther purification.

Step 6: 1-Benzenesulfonyl-4-morpholin-4-ylmethyl-1H-indole-2-carboxylicacid methyl ester

A mixture of1-benzenesulfonyl-4-morpholin-4-ylmethyl-1H-indole-2-carboxylic acid(670 mg, 1.67 mmol) and SOCl₂ (10 mL, 137 mmol) was heated at reflux for1 h. The solvent was evaporated under reduced pressure and the residuewas dried under vacuum for 1 h. THF (20 mL) was added, followed by MeOH(1.5 mL) and the reaction mixture was stirred at room temperature for 1h. The solved was evaporated under reduced pressure. The residue waspurified by chromatography (silica gel, 60% EtOAc/hexanes) to give1-benzenesulfonyl-4-morpholin-4-ylmethyl-1H-indole-2-carboxylic acidmethyl ester (486 mg, 70%) as an oil.

Step 7:3-(1-Benzenesulfonyl-4-morpholin-4-ylmethyl-1H-indol-2-yl)-3-oxo-propionitrile

A mixture of1-benzenesulfonyl-4-morpholin-4-ylmethyl-1H-indole-2-carboxylic acidmethyl ester (486 mg, 1.17 mmol), CH₃CN (0.37 mL, 7.0 mmol) and THF (10mL) was cooled to −78° C. LDA (2M in THF; 12 mL, 2.4 mmol) was added andthe mixture was stirred at −78° C. for 30 min. Saturated aqueous NH₄Clsolution (10 mL) and then water (50 mL) were added. The mixture wasextracted with EtOAc (350 mL). The organic layer was washed with brine,dried (Na₂SO₄), filtered, and evaporated to give3-(1-benzenesulfonyl-4-morpholin-4-ylmethyl-1H-indol-2-yl)-3-oxo-propionitrile(506 mg, 93%) as an oil which was used directly in the next step withfurther purification.

Step 8:(E)-2-(1-Benzenesulfonyl-4-morpholin-4-ylmethyl-1H-indole-2-carbonyl)-3-dimethylamino-acrylonitrile

A mixture of3-(1-benzenesulfonyl-4-morpholin-4-ylmethyl-1H-indol-2-yl)-3-oxo-propionitrile(506 mg, 1.17 mmol), N,N-dimethylformamide dimethyl acetal (0.21 mL,1.55 mmol), and toluene (10 mL) was stirred at room temperatureovernight. The solvent was evaporated. The residue was purified bychromatography (silica gel, 100% EtOAc) to give(E)-2-(1-benzenesulfonyl-4-morpholin-4-ylmethyl-1H-indole-2-carbonyl)-3-dimethylamino-acrylonitrile(285 mg, 50%) as a yellow foam.

Step 9:{5-Amino-1-[4-(2,3-difluoro-phenoxy)-2-methyl-phenyl]-1H-pyrazol-4-yl}-(1-benzenesulfonyl-4-morpholin-4-ylmethyl-1H-indol-2-yl)-methanone

A mixture of [4-(2,3-difluoro-phenoxy)-2-methyl-phenyl]-hydrazine (whichmay be prepared as described for Intermediate 1; 130 mg, 0.52 mmol),(E)-2-(1-benzenesulfonyl-4-morpholin-4-ylmethyl-1H-indole-2-carbonyl)-3-dimethylamino-acrylonitrile(60 mg, 0.125 mmol), and EtOH (10 mL) was heated at reflux overnight.The solvent was removed under reduced pressure. The residue was purifiedby chromatography (silica gel, 50% EtOAc/hexanes) to give{5-amino-1-[4-(2,3-difluoro-phenoxy)-2-methyl-phenyl]-1H-pyrazol-4-yl}-(1-benzenesulfonyl-4-morpholin-4-ylmethyl-1H-indol-2-yl)-methanone(56 mg, 65%) as an oil.

Step 10:{5-Amino-1-[4-(2,3-difluoro-phenoxy)-2-methyl-phenyl]-1H-pyrazol-4-yl}-(4-morpholin-4-ylmethyl-1H-indol-2-yl)-methanone

A mixture of{5-amino-1-[4-(2,3-difluoro-phenoxy)-2-methyl-phenyl]-1H-pyrazol-4-yl}-(1-benzenesulfonyl-4-morpholin-4-ylmethyl-1H-indol-2-yl)-methanone(56 mg, 0.082 mmol), Cs₂CO₃ (133 mg, 0.41 mmol), THF (4 mL) and MeOH (2mL) was stirred at room temperature overnight. The solvent was removedunder reduced pressure. The residue was purified by chromatography(silica gel, 70% EtOAc/hexanes) to give{5-amino-1-[4-(2,3-difluoro-phenoxy)-2-methyl-phenyl]-1H-pyrazol-4-yl}-(4-morpholin-4-ylmethyl-1H-indol-2-yl)-methanone(35 mg, 79%) as a light yellow oil. ¹H NMR (400 MHz, CDCl₃) δ ppm 9.27(br. s., 1H), 8.27 (s, 1H), 7.62 (s, 1H), 7.39 (d, J=8.0 Hz, 1H), 7.35(d, J=8.5 Hz, 1H), 7.30 (d, J=7.3 Hz, 1H), 3.86 (br. s., 2H), 3.75 (br.s., 4H), 2.54 (br. s., 3H), 2.20 (s, 3H). MS calcd. for C₃₀H₂₈F₂N₅O₃[(M+H)⁺] 544, obsd. 544.1.

Example I-203-{4-[5-Amino-4-(4-cyanomethyl-1H-indole-2-carbonyl)-pyrazol-1-yl]-3-methyl-phenoxy}-benzonitrile

Step 1:3-{4-[5-Amino-4-(1-benzenesulfonyl-4-cyanomethyl-1H-indole-2-carbonyl)-pyrazol-1-yl]-3-methyl-phenoxy}-benzonitrile

A mixture of3-{4-[5-amino-4-(1-benzenesulfonyl-4-bromo-1H-indole-2-carbonyl)-pyrazol-1-yl]-3-methyl-phenoxy}-benzonitrile(which may be prepared as described for Example I-14 Step 1; 107 mg,0.164 mmol), tert-butyl cyanoacetate (81 mg, 0.57 mmol),bis(tri-tert-butylphosphine)palladium(0) (Strem Chemicals; 29.5 mg,0.058 mmol), trisodium phosphate (121 mg, 0.74 mmol) and toluene (2 mL)was heated in a sealed tube at 100° C. overnight. Water (2 mL) was addedand the mixture was extracted with EtOAc (2×5 mL). The combined organiclayers were evaporated. The residue was purified by chromatography(silica gel, 40% EtOAc/hexanes) to give3-{4-[5-amino-4-(1-benzenesulfonyl-4-cyanomethyl-1H-indole-2-carbonyl)-pyrazol-1-yl]-3-methyl-phenoxy}-benzonitrile(34 mg, 34%) as an oil.

Step 2:3-{4-[5-Amino-4-(4-cyanomethyl-1H-indole-2-carbonyl)-pyrazol-1-yl]-3-methyl-phenoxy}-benzonitrile

A mixture of3-{4-[5-amino-4-(1-benzenesulfonyl-4-cyanomethyl-1H-indole-2-carbonyl)-pyrazol-1-yl]-3-methyl-phenoxy}-benzonitrile(34 mg, 0.056 mmol), Cs₂CO₃ (81.4 mg, 0.25 mmol), THF (4 mL) and MeOH (2mL) was stirred at room temperature overnight. The solvent was removedunder reduced pressure. The residue was purified by chromatography(silica gel, 40% EtOAc/hexanes) to give3-{4-[5-amino-4-(4-cyanomethyl-1H-indole-2-carbonyl)-pyrazol-1-yl]-3-methyl-phenoxy}-benzonitrile(11 mg, 42%) as a light yellow solid, ¹H NMR (400 MHz, CDCl₃) δ ppm 9.38(br. s., 1H), 8.31 (s, 1H), 7.46-7.58 (m, 4H), 7.31-7.45 (m, 7H),7.22-7.26 (m, 1H), 6.99-7.11 (m, 3H), 4.10 (s, 2H), 2.25 (s, 4H). MScalcd. for C₂₈H₂₁N₆O₂ [(M+H)⁺] 473, obsd. 473.

Example I-212-{5-Amino-1-[4-(3-cyano-phenoxy)-2-methyl-phenyl]-1H-pyrazole-4-carbonyl}-1H-indole-4-carboxylicacid methylamide trifluoroacetate salt

A sealed tube containing3-{4-[5-amino-4-(4-bromo-1H-indole-2-carbonyl)-pyrazol-1-yl]-3-methyl-phenoxy}-benzonitrile(which may be prepared as described in Example I-14 Step 2; 160 mg, 0.31mmol), Pd(PPh₃)₄ (108 mg, 0.094 mmol), methylamine (2 M THF; 2 mL, 4mmol) and THF (20 mL) was charged with 30 psi of carbon monoxide. Themixture was heated at 95° C. overnight and then the solvent wasevaporated. The residue was purified by preparative HPLC to give2-{5-amino-1-[4-(3-cyano-phenoxy)-2-methyl-phenyl]-1H-pyrazole-4-carbonyl}-1H-indole-4-carboxylicacid methylamide trifluoroacetate salt (73 mg, 48%) as an off-whitelyophilized powder. ¹H NMR (400 MHz, CDCl₃) δ ppm 9.43 (s, 1H), 8.36 (s,1H), 7.89 (dd, J=2.1, 0.9 Hz, 1H), 7.60-7.64 (m, 1H), 7.44-7.55 (m, 3H),7.38-7.44 (m, 2H), 7.31-7.37 (m, 2H), 7.07 (d, J=2.8 Hz, 1H), 7.01 (dd,J=8.4, 2.4 Hz, 1H), 3.15 (d, J=4.8 Hz, 3H), 2.22 (s, 3H). MS calcd. forC₂₈H₂₁F₂N₆O₂ [(M+H)⁺] 511, obsd. 511.

BIOLOGICAL EXAMPLES

Bruton's Tyrosine Kinase (Btk) Inhibition Assay

The assay is a capture of radioactive ³³P phosphorylated product throughfiltration. The interactions of Btk, biotinylated SH₂ peptide substrate(Sre homology), and ATP lead to phosphorylation of the peptidesubstrate. Biotinylated product is bound streptavidin sepharose beads.All bound, radiolabeled products are detected by scintillation counter.

Plates assayed are 96-well polypropylene (Greiner) and 96-well 1.2 μmhydrophilic PVDF filter plates (Millipore). Concentrations reported hereare final assay concentrations: 10-100 μM compounds in DMSO (Burdick andJackson), 5-10 nM Btk enzyme (His-tagged, full-length), 30 μM peptidesubstrate (Biotin-Aca-AAAEEIYGEI-NH₂), 100 μM ATP (Sigma), 8 mMimidazole (Sigma, pH 7.2), 8 mM glycerol-2-phosphate (Sigma), 200 μMEGTA (Roche Diagnostics), 1 mM MnCl₂ (Sigma), 20 mM MgCl₂ (Sigma), 0.1mg/ml BSA (Sigma), 2 mM DTT (Sigma), 1 μCi ³³P ATP (Amersham), 20%streptavidin sepharose heads (Amersham), 50 mM EDTA (Gibco), 2 M NaCl(Gibco), 2 M NaCl w/1% phosphoric acid (Gibco), microscint-20 (PerkinElmer).

IC₅₀ determinations are calculated from 10 data points per compoundutilizing data produced from a standard 96-well plate assay template.One control compound and seven unknown inhibitors were tested on eachplate and each plate was run twice. Typically, compounds were diluted inhalf-log starting at 100 μM and ending at 3 nM. The control compound wasstaurosporine. Background was counted in the absence of peptidesubstrate. Total activity was determined in the presence of peptidesubstrate. The following protocol was used to determine Btk inhibition.

1) Sample preparation: The test compounds were diluted at half-logincrements in assay buffer (imidazole, glycerol-2-phosphate, EGTA,MnCl₂, MgCl₂, BSA).

2) Bead preparation

-   -   a.) rinse beads by centrifuging at 500 g    -   b.) reconstitute the heads with PBS and EDTA to produce a 20%        bead slurry

3) Pre-incubate reaction mix without substrate (assay buffer, DTT, ATP,³³P ATP) and mix with substrate (assay buffer, DTT, ATP, ³³P ATP,peptide substrate) 30° C. for 15 min.

4) To start assay, pie-incubate 10 μL Btk in enzyme buffer (imidazole,glycerol-2-phosphate, BSA) and 10 μL of test compounds for 10 min at RT.

5) Add 30 μL reaction mixture without or with substrate to Btk andcompounds.

6) Incubate 50 μL total assay mix for 30 min at 30° C.

7) Transfer 40 μL of assay to 150 μL bead slurry in filter plate to stopreaction.

8) Wash filter plate after 30 min, with following steps

-   -   a. 3×250 μL NaCl    -   b. 3×250 μL NaCl containing 1% phosphoric acid    -   c. 1×250 μL H₂O

9) Dry plate for 1 h at 65° C. or overnight at RT

10) Add 50 μL microscint-20 and count ³³P cpm on scintillation counter.

-   -   Calculate percent activity from raw data in cpm        percent activity=(sample−bkg)/(total activity−bkg)×100    -   Calculate IC₅₀ from percent activity, using one-site dose        response sigmoidal model        y=A+((B−A)/(1+((x/C)^(D)))))    -   x=cmpd conc, y=% activity, A=min, B=max, C=IC₅₀, D=1 (hill        slope)

Bruton's Tyrosine Kinase (BTK) Inhibition TR-FRET (Time Resolved FRET)Assay

This BTK competition assay measures compound potency (IC50) for theinactivated state of Bruton's Tyrosine Kinase using FRET(Förster/Flouresence Resonance Energy Transfer) technology. The BTK—Eucomplex was incubated on ice one hour prior to use at a startingconcentration of 50 nM BTK-Bioease™: 10 nM Eu-streptavidin (Perkin-ElmerCatalog #AD0062). The assay buffer consisted of 20 mM HEPES (pH 7.15),0.1 mM DTT, 10 mM MgCl₂, 0.5 mg/ml BSA with 3% Kinase Stabilizer(Fremont Biosolutions, Catalog #STB-K02). After 1 h, the reactionmixture from above was diluted 10 fold in assay buffer to make 5 nM BTK:1 nM Eu-Streptavidin complex (donor fluorophore). 18 μl of a mixture of0.11 nM BTK-Eu and 0.11 nM Kinase Tracer 178 (Invitrogen, Catalog#PV5593,) with BTK-Eu alone as no negative control, was then dispensedinto 384-well flat bottom plates (Greiner, 784076). Compounds to betested in assay were prepared as 10× concentrations and serial dilutionin half-log increments was performed in DMSO so as to generate 10 pointcurves. To initiate the FRET reaction, compounds prepared as 10× stockin DMSO was added to the plates and the plates were incubated 18-24 h at14+ C.

After the incubation the plates were read on a BMG Pherastar Fluorescentplate reader (or equivalent) and used to measure the emission energyfrom the europium donor fluorophore (620 nm emission) and the FRET (665nm emission). The negative control well values were averaged to obtainthe mean minimum. The positive “no inhibitor” control wells wereaveraged to Obtain the mean maximum. Percent of maximal FRET wascalculated using following equation:% max FRET=100×[(FSR _(cmpd) −FSR _(mean min))/(FSR _(mean max) −FSR_(mean min))]

where FSR=FRET Signal ratio. % Max FRET curves were plotted in ActivityBase (Excel) and the IC50(%), hill slope, z′ and % CV were determined.The mean IC50 and standard deviation will be derived from duplicatecurves (singlet inhibition curves from two independent dilutions) usingMicrosoft Excel.

Representative compound data for this assay are listed below in TableII.

TABLE II FRET IC50 Compound (μmol) 1 0.06152 2 0.08512 3 0.10574 40.0208 5 0.2135 6 0.01189 7 0.06531 8 0.00615 9 0.03306 10 0.00421 110.05911 12 0.20117 13 0.02952 14 0.00838 15 0.09358 16 0.03905 170.02106 18 19 0.00559 20 0.00845 21 0.01302

Inhibition of B Cell Activation in Whole Blood Measured by CD69Expression

A procedure to test the ability of Btk inhibitors to suppress B cellreceptor-mediated activation of B cells in human blood is as follows:

Human whole blood (HWB) is obtained from healthy volunteers, with thefollowing restrictions; 24 hr drug-free, non-smokers. Blood is collectedby venipuncture into Vacutainer tubes anticoagulated with sodiumheparin. Test compounds are diluted to ten times the desired startingdrug concentration in PBS (20×), followed by three-fold serial dilutionsin 10% DMSO in PBS to produce a nine point dose-response curve. 5.5 μlof each compound dilution is added in duplicate to a 2 ml 96-well Vbottom plate (Analytical Sales and Services, #59623-23); 5.5 μl of 10%DMSO in PBS is added to control and no-stimulus wells. HWB (loop isadded to each well, and after mixing the plates are incubated at 37 C,5% CO₂. 100% humidity for 30 minutes. Goat F(ab′)2 anti-human IgM(Southern Biotech, #2022-14) (10 μl of a 500 μg/ml solution, 50 μg/mlfinal concentration) is added to each well (except the no-stimuluswells) with mixing and the plates are incubated for an additional 20hours.

At the end of the 20 hour incubation, samples are incubated withflorescent-probe-labeled antibodies (15 μl PE Mouse anti-Human CD20, BDPharmingen, #555623, and/or 20 μl APC Mouse anti-Human CD69, BDPharmingen #555533) for 30 minutes, at 37 C, 5% CO₂, 100% humidity.Included are induced control, unstained and single stains forcompensation adjustments and initial voltage settings. Samples are thenlysed with 1 ml of 1× Pharmingen Lyse Buffer (BD Pharmingen #555899),and plates are centrifuged at 1800 rpm for 5 minutes. Supernatants areremoved via suction and the remaining pellets are lysed again withanother 1 ml of 1× Pharmingen Lyse Buffer, and plates are spun down asbefore. Supernatants are aspirated and remaining pellets are washed inFACs buffer (PBS+1% FBS). After a final spin, the supernantants areremoved and pellets are resuspended in 180 μl of FACs buffer. Samplesare transferred to a 96 well plate suitable to be run on the HTS 96 wellsystem on the BD LSR II flow cytometer.

Using appropriate excitation and emission wavelengths for thefluorophores used, data are acquired and percent positive cell valuesare obtained using Cell Quest Software. Results are initially analyzedby FACS analysis software (Flow Jo). The IC50 for test compounds isdefined as the concentration which decreases by 50% the percentage ofCD69-positive cells that are also CD20-positive after stimulation byanti-IgM (average of 8 control wells, after subtraction of the averageof 8 wells for the no-stimulus background). The IC50 values arecalculated using XLfit software version 3, equation 201.

Inhibition of B-Cell Activation—B Cell FLIPR Assay in Ramos Cells

Inhibition of B-cell activation by compounds of the present invention isdemonstrated by determining the effect of the test compounds on anti-IgMstimulated B cell responses.

The B cell FLIPR assay is a cell based functional method of determiningthe effect of potential inhibitors of the intracellular calcium increasefrom stimulation by an anti-IgM antibody. Ramos cells (human Burkitt'slymphoma cell line. ATCC-No. CRL-1596) were cultivated in Growth Media(described below). One day prior to assay, Ramos cells were resuspendedin fresh growth media (same as above) and set at a concentration of0.5×10⁶/mL in tissue culture flasks. On day of assay, cells are countedand set at a concentration of 1×10⁶/mLl in growth media supplementedwith 1 M FLUO-3AM(TefLabs Cat-No. 0116, prepared in anhydrous DMSO and10% Pluronic acid) in a tissue culture flask, and incubated at 37° C.(4% CO₂) for one h. To remove extracellular dye, cells were collected bycentrifugation (5 min. 1000 rpm), resuspended in FLIPR buffer (describedbelow) at 1×10⁶ cells/mL and then dispensed into 96-well poly-D-lysinecoated black clear plates (BD Cat-No. 356692) at 1×10⁵ cells per well.Test compounds were added at various concentrations ranging from 100 μMto 0.03 μM (7 concentrations, details below), and allowed to incubatewith cells for 30 min at RT. Ramos cell Ca²⁺ signaling was stimulated bythe addition of 10 g/mL anti-IgM (Southern Biotech, Cat-No. 2020-01) andmeasured on a FLIPR (Molecular Devices, captures images of 96 wellplates using a CCD camera with an argon laser at 480 nM excitation).

Media/Buffers:

Growth Medium: RPMI 1640 medium with L-glutamine (Invitrogen, Cat-No.61870-010), 10% Fetal Bovine Serum (FBS, Summit Biotechnology Cat-No.FP-100-05): 1 mM Sodium Pyruvate (Invitrogen Cat. No. 11360-070).

FLIPR buffer: HBSS (Invitrogen, Cat-No. 141175-079), 2 mM CaCl₂ (SigmaCat-No. C-4901), HEPES (Invitrogen, Cat-No. 15630-080), 2.5 mMProbenecid (Sigma, Cat-No. P-8761), 0.1% BSA (Sigma, Cat-No. A-7906), 11mM Glucose (Sigma, Cat-No. G-7528)

Compound Dilution Details:

In order to achieve the highest final assay concentration of 100 μM, 24μL of 10 mM compound stock solution (made in DMSO) is added directly to576 μL of FLIPR buffer. The test compounds are diluted in FLIPR Buffer(using Biomek 2000 robotic pipettor) resulting in the following dilutionscheme: vehicle, 1.00×10⁻⁴ M, 1.00×10⁻⁵, 3.16×10⁻⁶, 1.00×10⁻⁶,3.16×10⁻⁷, 1.00×10⁻⁷, 3.16×10⁻⁸.

Assay and Analysis:

Intracellular increases in calcium were reported using a max-minstatistic (subtracting the resting baseline from the peak caused byaddition of the stimulatory antibody using a Molecular Devices FLIPRcontrol and statistic exporting software. The IC₅₀ was determined usinga non-linear curve fit (GraphPad Prism software).

Mouse Collagen-Induced Arthritis (mCIA).

On day 0 mice are injected at the base of the tail or several spots onthe back with an emulsion of Type II Collagen (i.d.) in CompleteFreund's adjuvant (CFA). Following collagen immunization, animals willdevelop arthritis at around 21 to 35 days. The onset of arthritis issynchronized (boosted) by systemic administration of collagen inIncomplete Freund's adjuvant (IFA: i.d.) at day 21. Animals are examineddaily after day 20 for any onset of mild arthritis (score of 1 or 2; seescore description below) which is the signal to boost. Following boost,mice are scored and dosed with candidate therapeutic agents for theprescribed time (typically 2-3 weeks) and dosing frequency, daily (QD)or twice-daily (BID).

Rat Collagen-Induced Arthritis (rCIA)

On day 0, rats are injected with an emulsion of Bovine Type II Collagenin Incomplete Freund's adjuvant (IFA) is injected intradermally (i.d.)on several locations on the back. A booster injection of collagenemulsion is given around day 7. (i.d.) at the base of the tail oralternative sites on the back. Arthritis is generally observed 12-14days after the initial collagen injection. Animals may be evaluated forthe development of arthritis as described below (Evaluation ofarthritis) from day 14 onwards. Animals are dosed with candidatetherapeutic agents in a preventive fashion starting at the time ofsecondary challenge and for the prescribed time (typically 2-3 weeks)and dosing frequency, daily (QD) or twice-daily (BID).

Evaluation of Arthritis:

In both models, developing inflammation of the paws and limb joints isquantified using a scoring system that involves the assessment of the 4paws following the criteria described below:

Scoring: 1=swelling and/or redness of paw or one digit.

-   -   2=swelling in two or more joints.    -   3=gross swelling of the paw with more than two joints involved.    -   4=severe arthritis of the entire paw and digits.

Evaluations are made on day 0 for baseline measurement and startingagain at the first signs or swelling for up to three times per weekuntil the end of the experiment. The arthritic index for each mouse isobtained by adding the four scores of the individual paws, giving amaximum score of 16 per animal.

Rat In Vivo Asthma Model

Male Brown-Norway rats are sensitized i.p. with 100 μg of OA (ovalbumin)in 0.2 ml alum once every week for three weeks (day 0, 7, and 14). Onday 21 (one week following last sensitization), the rats are dosed q.d.with either vehicle or compound formulation subcutaneously 0.5 hourbefore OA aerosol challenge (1% OA for 45 minutes) and terminated 4 or24 hours after challenge. At time of sacrifice, serum and plasma arecollected from all animals for serology and PK, respectively. A trachealcannula is inserted and the lungs are lavaged 3× with PBS. The BAL fluidis analyzed for total leukocyte number and differential leukocytecounts. Total leukocyte number in an aliquot of the cells (20-100 μl) isdetermined by Coulter Counter. For differential leukocyte counts, 50-200μl of the sample is centrifuged in a Cytospin and the slide stained withDiff-Quik. The proportions of monocytes, eosinophils, neutrophils andlymphocytes are counted under light microscopy using standardmorphological criteria and expressed as a percentage. Representativeinhibitors of Btk show decreased total leucocyte count in the BAL of OAsensitized and challenged rats as compared to control levels.

The foregoing invention has been described in some detail by way ofillustration and example, for purposes of clarity and understanding. Itwill be obvious to one of skill in the art that changes andmodifications may be practiced within the scope of the appended claims.Therefore, it is to be understood that the above description is intendedto be illustrative and not restrictive. The scope of the inventionshould, therefore, be determined not with reference to the abovedescription, but should instead be determined with reference to thefollowing appended claims, along with the full scope of equivalents towhich such claims are entitled.

All patents, patent applications and publications cited in thisapplication are hereby incorporated by reference in their entirety forall purposes to the same extent as if each individual patent, patentapplication or publication were so individually denoted.

The invention claimed is:
 1. A method for treating an inflammatoryand/or autoimmune condition, comprising a step of administering to apatient in need thereof a therapeutically effective amount of a compoundof Formula I:

wherein: R¹ is H or halo; R² is H, halo, or cyano; R³ is R⁴ or R⁵; R⁴ ishalo or cyano; R⁵ is phenyl, heteroaryl, —C(═O)R^(5′), lower alkyl, orbenzyl, optionally substituted with one or more R^(5′); R^(5′) is loweralkyl, cyano, hydroxyl, heterocycloalkyl, phenyl, amino, alkyl amino,dialkyl amino, or lower alkoxy; and X is lower alkyl or halo; or apharmaceutically acceptable salt thereof.
 2. A method for treating aninflammatory condition, comprising a step of administering to a patientin need thereof a therapeutically effective amount of a compound ofFormula I:

wherein: R¹ is H or halo; R² is H, halo, or cyano; R³ is R⁴ or R⁵; R⁴ ishalo or cyano; R⁵ is phenyl, heteroaryl, —C(═O)R^(5′), lower alkyl, orbenzyl, optionally substituted with one or more R^(5′); R^(5′) is loweralkyl, cyano, hydroxyl, heterocycloalkyl, phenyl, amino, alkyl amino,dialkyl amino, or lower alkoxy; and X is lower alkyl or halo; or apharmaceutically acceptable salt thereof.